WO2023241618A1 - Aminopyrimidine compounds and use thereof - Google Patents

Aminopyrimidine compounds and use thereof Download PDF

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Publication number
WO2023241618A1
WO2023241618A1 PCT/CN2023/100166 CN2023100166W WO2023241618A1 WO 2023241618 A1 WO2023241618 A1 WO 2023241618A1 CN 2023100166 W CN2023100166 W CN 2023100166W WO 2023241618 A1 WO2023241618 A1 WO 2023241618A1
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compound
add
μmol
ethyl acetate
reaction
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PCT/CN2023/100166
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French (fr)
Chinese (zh)
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陈正霞
戴美碧
张杨
陈曙辉
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南京明德新药研发有限公司
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Publication of WO2023241618A1 publication Critical patent/WO2023241618A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D419/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms
    • C07D419/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • the present invention relates to a series of aminopyrimidine compounds and their applications, specifically to the compounds represented by formula (XI), their stereoisomers and their pharmaceutically acceptable salts and their applications.
  • EGFR Epidermal Growth Factor Receptor, referred to as EGFR, ErbB-1 or HER1
  • EGFR is a member of the epidermal growth factor receptor (HER) family.
  • EGFR is widely distributed on the surface of mammalian epithelial cells, fibroblasts, glial cells, keratinocytes and other cells.
  • the EGFR signaling pathway plays an important role in physiological processes such as cell growth, proliferation and differentiation. Mutated EGFR exists in many tumors, and many EGFR mutant types have been discovered.
  • EGFR mutations More than 80% of EGFR mutations are 19Del and L858R mutations, and the rest are rare mutations, mainly including G719X, E709X, and Del18 in exon 18, Ins19 in exon 19, and Ins20 in exon 20. , S768I, L861Q on exon 21.
  • Targeted therapy has shown excellent efficacy for patients with driver mutations in genes such as EGFR.
  • first-generation EGFR-TKIs such as gefitinib and erlotinib
  • first-generation EGFR-TKIs use competitive non-covalent binding to ATP to inhibit EGFR with deletion mutations in exon 19 and point mutations in exon 21.
  • first-generation EGFR-TKIs can also inhibit the EGFR function of normal cells, leading to side effects such as rash, diarrhea, and loss of appetite.
  • Second-generation EGFR-TKIs such as afatinib and dacomitinib, competitively and irreversibly bind to ATP through covalent bonding.
  • the inhibitory effect is stronger and the progression-free survival period mPFS is improved.
  • the disadvantage is that the side effects are stronger due to irreversible inhibition.
  • the T790M mutation occurs in the amino acid fragment bound to EGFR-TKIs, leading to drug resistance, accounting for about 50-70%.
  • the third-generation EGFR-TKIs AstraZeneca's AZD9291 (osimertinib) has become the first inhibitor to target the EGFRT790M mutation, targeting EGFR gene mutations in non-small cell lung cancer (including Exon18, 19, and 21 mutations). After 10 months of medication, the exon 20C797 mutation will occur, resulting in resistance to the third-generation drug AZD9291, and insufficient efficacy against the EGFR del19/L858R T790M C797S triple mutation. Therefore, the development of drugs that are selective for EGFR del19/L858R T790M C797S triple mutations while also targeting wild-type EGFR has huge clinical and market value.
  • the compound of the present invention has excellent inhibitory activity against the EGFR del19/L858R T790M C797S triple mutation, and at the same time has good selectivity for wild-type EGFT, and is expected to be used in patients with EGFR abnormal diseases.
  • the present invention provides the compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
  • T 1 and T 2 are independently selected from CH and N;
  • T 4 is selected from C and N;
  • T 5 is selected from CH and N;
  • Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
  • Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
  • R 2 is selected from H, halogen and OH, or is absent;
  • T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
  • R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
  • R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
  • R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
  • R 6 and R 7 form a double bond, making the structural unit form
  • R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 8 is selected from H and halogen
  • T 3 is selected from C(R c ) 2 , NR c and O;
  • n 0, 1, 2 and 3;
  • n is selected from 1, 2 and 3;
  • r is selected from 1, 2 and 3;
  • s is selected from 1 and 2;
  • t is selected from 1 and 2;
  • Each R a is independently selected from H and halogen
  • Each R b is independently selected from H and halogen
  • R c is selected from H and C 1-3 alkyl
  • ring A is selected from 2,3-dihydrobenzofuryl, phthalyl, 2,3 -Dihydropyridofuranyl and isoindolinyl;
  • Ring B is not selected from pyrazolyl.
  • the present invention also provides the compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
  • T 1 and T 2 are independently selected from CH and N;
  • T 4 is selected from C and N;
  • T 5 is selected from CH and N;
  • Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
  • Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
  • the C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
  • R 2 is selected from H, halogen and OH, or is absent;
  • T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
  • R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
  • R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
  • R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
  • R 6 and R 7 form a double bond, making the structural unit form
  • R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 8 is selected from H and halogen
  • T 3 is selected from CR c , NR c and O;
  • n 0, 1, 2 and 3;
  • n is selected from 1, 2 and 3;
  • r is selected from 1, 2 and 3;
  • s is selected from 1 and 2;
  • t is selected from 1 and 2;
  • Each R a is independently selected from H and halogen
  • Each R b is independently selected from H and halogen
  • R c is selected from H and C 1-3 alkyl
  • the present invention also provides the compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
  • T 1 and T 2 are independently selected from CH and N;
  • T 4 is selected from C and N;
  • T 5 is selected from CH and N;
  • Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
  • Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
  • R 2 is selected from H, halogen and OH, or is absent;
  • T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
  • R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
  • R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
  • R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
  • R 6 and R 7 form a double bond, making the structural unit form
  • R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 8 is selected from H and halogen
  • T 3 is selected from CR c , NR c and O;
  • n 0, 1, 2 and 3;
  • n is selected from 1, 2 and 3;
  • r is selected from 1, 2 and 3;
  • s is selected from 1 and 2;
  • t is selected from 1 and 2;
  • Each R a is independently selected from H and halogen
  • Each R b is independently selected from H and halogen
  • R c is selected from H and C 1-3 alkyl
  • ring A is selected from 2,3-dihydrobenzofuryl, phthalyl, 2,3 -Dihydropyridofuranyl and isoindolinyl;
  • Ring B is not selected from pyrazolyl.
  • the present invention also provides the compound represented by formula (V), its stereoisomer or its pharmaceutically acceptable salt,
  • T 1 and T 2 are independently selected from CH and N;
  • T 4 is selected from C and N;
  • T 5 is selected from CH and N;
  • Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
  • Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
  • the C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
  • T 3 is selected from NR c and O;
  • R 2 is selected from H, halogen and OH, or is absent;
  • T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
  • R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
  • R 4 is selected from F, OH, CH 3 , OCH 3 , CH 2 CH 3 and N(CH 3 ) 2 ;
  • R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
  • R 6 and R 7 form a double bond, making the structural unit form
  • R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group
  • n 0, 1, 2 and 3;
  • n is selected from 1, 2 and 3;
  • r is selected from 1, 2 and 3;
  • s is selected from 1 and 2;
  • t is selected from 1 and 2;
  • Each R a is independently selected from H and halogen
  • Each R b is independently selected from H and halogen
  • R c is selected from H and C 1-3 alkyl
  • R b is selected from H
  • T 1 is selected from N
  • T 2 is selected from N
  • T 4 is selected from C
  • the structural unit Selected from Where T 7 and T 8 are independently selected from N and CR
  • R is selected from H, halogen and CH 3 ,
  • Ring B is selected from pyrazolyltetrahydropyridyl, dihydropyranyl, cyclohexyl and cyclohexenyl, or;
  • R 2 is selected from H and R 6 is selected from H, R 7 is not H and not CH 3 , or;
  • R 6 is not H and is not CH 3 .
  • the present invention provides the compound represented by formula (V), its stereoisomer or its pharmaceutically acceptable salt,
  • T 1 and T 2 are independently selected from CH and N;
  • T 4 is selected from C and N;
  • T 5 is selected from CH and N;
  • Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl, and isoindoline;
  • Ring B is selected from pyrazolyl and piperidinyl
  • the C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
  • T 3 is selected from NR c and O;
  • R 2 is selected from H, halogen and OH, or is absent;
  • T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
  • R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
  • R 4 is selected from F, OH, CH 3 , OCH 3 and
  • R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
  • R 6 and R 7 form a double bond, making the structural unit form
  • R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group
  • n is selected from 1, 2 and 3;
  • n is selected from 1, 2 and 3;
  • r is selected from 1, 2 and 3;
  • s is selected from 1 and 2;
  • t is selected from 1 and 2;
  • Each R a is independently selected from H and halogen
  • Each R b is independently selected from H and halogen
  • R c is selected from H and C 1-3 alkyl
  • T 1 is selected from N
  • T 2 is selected from N
  • T 4 is selected from C
  • the structural unit Selected from Where T 7 and T 8 are independently selected from N and CR
  • R is selected from H, halogen and CH 3 ,
  • Ring B is selected from pyrazolyl, or
  • R 2 is selected from H and R 6 is selected from H, R 7 is not H and not CH 3 , or;
  • R 6 is not H and is not CH 3 .
  • the present invention also provides the compound represented by formula (V), its stereoisomer or its pharmaceutically acceptable salt,
  • T 1 and T 2 are independently selected from CH and N;
  • T 4 is selected from C and N;
  • T 5 is selected from CH and N;
  • Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- Dihydrobenzofuryl, phthalyl and 2,3-dihydropyridofuranyl;
  • Ring B is selected from pyrazolyl and piperidinyl
  • the C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
  • T 3 is selected from NR c and O;
  • R 2 is selected from H, halogen and OH, or is absent;
  • T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
  • R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group
  • R 3 is selected from halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optional Substituted by 1, 2 or 3 halogens;
  • R 4 is selected from F, OH, CH 3 , OCH 3 and
  • R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
  • R 6 and R 7 form a double bond, making the structural unit form
  • R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group
  • n is selected from 1, 2 and 3;
  • n is selected from 1, 2 and 3;
  • s is selected from 1 and 2;
  • t is selected from 1 and 2;
  • Each R a is independently selected from H and halogen
  • Each R b is independently selected from H and halogen
  • R c is selected from H and CH 3 ;
  • Ring B is selected from pyrazolyl, or
  • R 2 , R 6 , R 7 and R a are not selected from H at the same time, or
  • T 2 is selected from CH.
  • T 3 is selected from CH 2 , NH, NCH 3 , NCH 2 CH 3 , NCH(CH 3 ) 2 and O, and other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • R 2 is selected from H, F and OH, or does not exist, and other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • R 6 and R 7 are independently selected from H and CH 3 , and other variables are as defined in the present invention.
  • R 6 is selected from H and CH 3 , and other variables are as defined in the present invention.
  • R 7 is selected from H and CH 3 , and other variables are as defined in the present invention.
  • R 8 is selected from H and F, and other variables are as defined in the present invention.
  • R 3 is selected from F, CH 3 , CH 2 CH 3 , CH(CH 3 ) 2 , OCH 3 and cyclopropyl, and other variables are as defined in the present invention.
  • R 3 is selected from F, CH 3 , CH(CH 3 ) 2 , OCH 3 and cyclopropyl, and other variables are as defined in the present invention.
  • the above-mentioned Ring A is selected from 2,3-dihydrobenzofuranyl, phenyl and pyridyl, and other variables are as defined in the present invention.
  • the above-mentioned ring A is selected from phenyl and pyridyl, and other variables are as defined in the present invention.
  • the above-mentioned ring B is selected from Other variables are as defined in the present invention.
  • the above-mentioned ring B is selected from Other variables are as defined in the present invention.
  • the above-mentioned ring B is selected from piperidyl, tetrahydropyridyl, dihydropyranyl, cyclohexenyl and 7-10 membered heterocycloalkyl, and other variables are as defined in the invention.
  • R 4 is selected from F, OH, CH 3 , OCH 3 , CH 2 CH 3 , N(CH 3 ) 2 and Other variables are as defined in the present invention.
  • the above compounds, their stereoisomers or their pharmaceutically acceptable salts are selected from,
  • R 1 is selected from
  • Ring A, Ring B, R2 , R3 , R4, R6 , R7 , R8 , T1 , T2 , T3 , T4 , T5 , m, n and r are as defined herein.
  • the above compounds, their stereoisomers or their pharmaceutically acceptable salts are selected from,
  • R 1 is selected from Rings A, R2 , R3 , R4 , R6 , R7 , T3 , m, n and r are as defined herein.
  • the present invention provides the following compounds, their stereoisomers or their pharmaceutically acceptable salts,
  • the compound, its stereoisomer or its pharmaceutically acceptable salt, the compound thereof is selected from,
  • the compound, its stereoisomer or its pharmaceutically acceptable salt, the compound thereof is selected from,
  • the present invention also provides a pharmaceutical composition, including a therapeutically effective amount of a compound as defined in the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier and diluent. or excipients.
  • the present invention also provides the use of the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt, or the above-mentioned composition in the preparation of drugs for treating solid tumors.
  • the solid tumor refers to solid tumors such as non-small cell lung cancer.
  • the present invention also provides the following synthesis methods of the above compounds:
  • the present invention also provides the following test methods for the above compounds:
  • Test method 1 Kinase inhibitory activity of compounds against EGFR (T790M/C797S/L858R)
  • reaction buffer (20mM hydroxyethyl piperazine ethyl sulfonic acid (Hepes) (pH 7.5), 10mM magnesium chloride (MgCl 2 ), 1mM ethylene glycol bisaminoethyl ether tetraacetic acid (EGTA), 0.02% polyoxyethylene ten Dialkyl ether (Brij35), 0.02mg/mL BSA, 0.1mM sodium vanadate (Na 3 VO 4 ), 2mM dithiothreitol (DTT), 1% DMSO) and then add a certain concentration of substrate and coenzyme factor , kinase and test compound (10 doses, 3-fold serial dilution, 2% DMSO final concentration) and mix well, incubate the mixture at room temperature for 20 minutes, add a certain concentration of 33 P-ATP to the reaction mixture to start the reaction , followed by incubation at room temperature for 120 minutes.
  • Hepes hydroxyethyl piperazine
  • the radioactivity of the reactants is detected by filtration-binding method.
  • the final kinase activity is expressed as the ratio of the remaining kinase activity in the test sample to the kinase activity in the DMSO control group.
  • the dose-response relationship curve was fitted and IC 50 was calculated using GraphPad software.
  • Test method 2 Inhibitory activity of compounds on Ba/F3-FL-EGFR (T790M/C797S/L858R) cell proliferation
  • Adenosine Tri-Phosphate is a common energy carrier in various life activities in nature and is the smallest unit of energy storage and transfer.
  • CellTiter-Glo TM live cell detection kit uses luciferase as the detection substance, and luciferase requires the participation of ATP during the luminescence process.
  • the cell line was Ba/F3-FL-EGFR (T790M/C797S/L858R) stably transfected cell line.
  • the cell lines were cultured in an incubator with culture conditions of 37°C and 5% CO2 . Passage regularly and use cells in the logarithmic growth phase for plating.
  • Inhibition Rate(Inh%) 100-(RLUDrug-RLUMin)/(RLUMax-RLUMin)*100%.
  • the term "pharmaceutically acceptable” refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue. , without undue toxicity, irritation, allergic reactions, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • salts refers to salts of compounds of the present invention prepared from compounds having specific substituents found in the present invention and relatively non-toxic acids or bases.
  • base addition salts can be obtained by contacting such compounds with a sufficient amount of base in pure solution or in a suitable inert solvent.
  • acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent.
  • Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base or acid addition salts.
  • the pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid groups or bases.
  • such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereoisomers isomer, the (D)-isomer, the (L)-isomer, as well as their racemic mixtures and other mixtures, such as enantiomeric or diastereomerically enriched mixtures, all of which belong to the present invention. within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
  • enantiomers or “optical isomers” refer to stereoisomers that are mirror images of each other.
  • cis-trans isomers or “geometric isomers” refers to the inability of the double bonds or single bonds of the carbon atoms in the ring to rotate freely.
  • diastereomer refers to stereoisomers whose molecules have two or more chiral centers and are in a non-mirror image relationship between the molecules.
  • use wedge-shaped solid line keys and wedge-shaped dotted keys Represents the absolute configuration of a three-dimensional center
  • using straight solid line keys and straight dotted keys Represent the relative configuration of the three-dimensional center with a wavy line
  • wedge-shaped solid line key or wedge-shaped dotted key or use tilde Represents a straight solid line key and straight dotted keys like generation surface like represent mixture.
  • tautomer or “tautomeric form” means that at room temperature, isomers with different functional groups are in dynamic equilibrium and can quickly convert into each other. If tautomers are possible (eg in solution), a chemical equilibrium of tautomers can be achieved.
  • proton tautomers also called proton transfer tautomers
  • proton migration tautomers include interconversions by proton migration, such as keto-enol isomerization and imine-enol isomerization. Amine isomerization.
  • Valence tautomers include interconversions through the reorganization of some bonding electrons.
  • keto-enol tautomerization is the tautomerization between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
  • the terms “enriched in an isomer,” “enantiomerically enriched,” “enriched in an enantiomer,” or “enantiomerically enriched” refer to one of the isomers or enantiomers.
  • the content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
  • isomeric excess or “enantiomeric excess” refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, then the isomer or enantiomeric excess (ee value) is 80% .
  • optically active (R)- and (S)-isomers as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliaries, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure desired enantiomer.
  • a diastereomeric salt is formed with a suitable optically active acid or base, and then the salt is formed by conventional methods known in the art. Diastereomeric resolution is performed and the pure enantiomers are recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using chiral stationary phases, optionally combined with chemical derivatization methods (e.g., generation of amino groups from amines). formate).
  • the compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound.
  • compounds can be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or C-14 ( 14 C).
  • deuterated drugs can be replaced by heavy hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs can reduce side effects and increase drug stability. , enhance efficacy, extend drug biological half-life and other advantages. All variations in the isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
  • substituted means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the specific atom is normal and the substituted compound is stable.
  • any variable e.g., R
  • its definition in each instance is independent.
  • said group may optionally be substituted by up to two R's, with independent options for R in each case.
  • substituents and/or variants thereof are permitted only if such combinations result in stable compounds.
  • linking group When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
  • any one or more sites of the group can be connected to other groups through chemical bonds.
  • connection mode of the chemical bond is non-positioned and there are H atoms at the connectable site, when the chemical bond is connected, the number of H atoms at the site will be reduced correspondingly with the number of connected chemical bonds and become the corresponding valence. group.
  • the chemical bond connecting the site to other groups can be a straight solid line bond straight dashed key or wavy lines express.
  • the straight solid line bond in -OCH 3 means that it is connected to other groups through the oxygen atom in the group;
  • the straight dotted bond in means that it is connected to other groups through both ends of the nitrogen atoms in the group;
  • the wavy lines in indicate that the phenyl group is connected to other groups through the 1 and 2 carbon atoms in the phenyl group;
  • C 1-3 alkyl is used to mean a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms.
  • the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) .
  • Examples of C 1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.
  • halogen or halogen by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
  • C 1-3 alkoxy means those alkyl groups containing 1 to 3 carbon atoms that are attached to the remainder of the molecule through an oxygen atom.
  • the C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy groups, etc.
  • Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
  • a heteroatom may occupy the attachment position of the heterocycloalkyl to the rest of the molecule.
  • the 7-10-membered heterocycloalkyl group includes 7-10-membered, 7-9-membered, 7-8-membered, 8-10-membered, 8-9-membered, 8-membered, 9-membered and 10-membered heterocycloalkyl groups, etc. Examples of 7-10 membered heterocycloalkyl groups include, but are not limited to wait.
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and methods well known to those skilled in the art. Equivalent replacement method, preferred Implementations include, but are not limited to, embodiments of the present invention.
  • the structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art.
  • single crystal X-ray diffraction uses a Bruker D8 venture diffractometer to collect diffraction intensity data on the cultured single crystal.
  • the light source is CuK ⁇ radiation, and the scanning mode is: ⁇ / ⁇ scanning. After collecting relevant data, the direct method is further used. (Shelxs97)
  • the absolute configuration can be confirmed by analyzing the crystal structure.
  • TMSOTf represents trimethylsilyl triflate
  • NaBH 4 represents sodium borohydride
  • Cs 2 CO 3 represents cesium carbonate
  • NBS represents bromosuccinimide
  • PE represents petroleum ether
  • EtOAc represents ethyl acetate
  • DCM dichloromethane
  • MeOH represents methanol
  • EtOH represents ethanol
  • TFA trifluoroacetic acid
  • Pd(dppf)Cl 2 represents 1,1-bis(diphenylphosphorus)ferrocene chloride Palladium
  • K 2 CO 3 represents potassium carbonate
  • PtO 2 represents platinum dioxide
  • NaH represents sodium hydrogen
  • THF represents tetrahydrofuran
  • Boc represents tert-butyl
  • DMF represents N,N-dimethylformamide
  • KOH represents potassium hydroxide
  • Xantphos-Pd-G4 represents palladium methanesulfonate [4,5-bisdiphenylphosphin
  • the compound of the present invention exhibits high kinase inhibitory activity against del19/L858R T790M C797S mutated EGFR, and is highly selective compared with the wild type. It has good proliferation inhibitory activity against EGFR triple mutant cells and has good proliferation inhibitory activity against EGFR wild type cells. Significantly weakened, with better selectivity.
  • the compound of the invention has good in vivo metabolic stability, excellent oral absorption drug exposure and good oral absorption bioavailability, and has good stability in human and mouse liver microsomes.
  • Figure 10 Binding mode diagram of compound J and EGFR mutant protein.
  • the molecular docking process was performed using Maestro ( Performed with Glide SP[1] in version 2020-1) and default options.
  • the co-crystal structure of mutant EGFR (PDB ID: 5HCY) was selected as the docking template.
  • hydrogen atoms were added using the Protein Preparation Wizard module of Maestro [2] and the OPLS3e force field was used.
  • the LigPrep module was used to generate the three-dimensional structure of the molecule and energy minimization was performed [3], and the ConfGen module was used to search for small molecule conformations [4].
  • Receptor Grid Generation module in Glide to generate the grid file required for docking, with the ligand in the crystal structure as the center of the docking box. Analyze the interaction type between the protein receptor and the ligand, and then select molecules with high potential for synthesis testing based on the calculated docking score and binding mode selection.
  • the compound of the present invention binds well to EGFR mutant protein.
  • reaction solution was concentrated under reduced pressure, and subjected to preparative HPLC chromatography (chromatographic column: Phenomenex luna C18 250 ⁇ 80mm ⁇ 10 ⁇ m; mobile phase: [H 2 O (NH 3 H 2 O+NH 4 HCO 3 )-acetonitrile]; acetonitrile %: 0%-20%, 20min) to separate the product for 1 hour, and then separate it through chiral preparation (chromatographic column: DAICEL CHIRALCEL OJ (250mm ⁇ 50mm, 10 ⁇ m); mobile phase: supercritical CO 2 - [ethanol solution containing 0.1% ammonia] ; Containing 0.1% ammonia in ethanol solution%: 40%-40%) to obtain compounds 1h-1, 1h-2, 1h-3 and 1h-4.
  • Potassium monopersulfate (8.91g, 14.49mmol) was added to a solution of compound 1a (2.1g, 9.66mmol) in THF (20mL)/EtOH (20mL)/H 2 O (20mL), and the mixture was stirred at 25°C for 2 hours.
  • Methyl Grignard reagent (3.32g, 27.88mmol) was added dropwise to N-Boc-azetidine-3-carboxylic acid methyl ester (2.0g, 9.29mmol) in THF (25mL) at 0°C, and the temperature was raised to room temperature. , reaction 16hr. Add saturated ammonium chloride solution dropwise to quench the reaction, extract with 3 ⁇ 15 mL ethyl acetate, combine the organic layers, and concentrate under reduced pressure to obtain intermediate 2a.
  • 1 H NMR 400MHz, DMSO-d 6 ) ⁇ 3.72 (br s, 4H) 2.38-2.47 (m, 1H) 1.36 (s, 9H) 0.99 (s, 6H).
  • NBS (257.23 mg, 1.45 mmol) was added to the acetonitrile (5 mL) solution containing intermediate 2c (0.4 g, 1.45 mmol), and the reaction was carried out at 25°C for 2 hours.
  • Add 2 mL of water to the reaction solution to quench the reaction, evaporate the organic solvent under reduced pressure, extract with 2 ⁇ 10 mL of ethyl acetate, dry and concentrate the organic phase and perform silica gel column chromatography (DCM:MeOH 100:1) to obtain compound 2d.
  • MS m/z:356.8[M+3] + MS m/z:356.8[M+3] + .
  • Triethylamine (19.05g, 188.24mmol, 26.20mL) was added to a solution of compound 5a (30g, 125.36mmol) in DCM (300mL), cooled to -20°C, and then methanesulfonyl chloride (17.76g, 155.04mmol) , 12.00 mL) was added dropwise to the reaction solution, raised to 0°C and stirred for 1 hour.
  • the reaction solution was slowly poured into ice water (300 mL), the organic phase was collected in layers, and then washed with 10% sodium bicarbonate solution. Dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain compound 5b.
  • Diethyl malonate (28.86g, 180.21mmol, 27.23mL) was added dropwise to a solution of sodium hydrogen (6.54g, 163.50mmol, 60%) in N,N-dimethylformamide (180mL) at 0°C. Stir below 25°C for 1 hour, then add compound 5b (26g, 81.92mmol) in N,N-dimethylformamide (40mL), and stir at 70°C for 16 hours. Add the reaction liquid to ice water (400mL), add ethyl acetate for extraction (200mL ⁇ 3), combine the organic phases, wash with water (300mL ⁇ 3), and extract with saturated sodium chloride aqueous solution (300mL).
  • Trifluoroacetic acid (775.75 mg, 6.80 mmol, 503.73 ⁇ L) was added to a solution of compound 5h (0.1 g, 382.65 ⁇ mol) in DCM (2 mL) at 0°C, and the mixture was stirred at 25°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 5i.
  • 1 H NMR 400MHz, CDCl 3 ) ⁇ 7.81-9.30 (m, 1H), 4.31 (br s, 4H), 3.62-4.08 (m, 4H), 2.92-3.55 (m, 2H).
  • the crude compound 7 was separated by preparative high performance liquid chromatography (chromatographic column: Phenomenex C18 75 ⁇ 30mm ⁇ 3 ⁇ m; mobile phase: A (acetonitrile) and B (water, containing 0.05% ammonia water and 0.01mol ammonium bicarbonate); gradient: B% :5%-45%) to obtain compound 7.
  • methanol methanol
  • n-heptane n-heptane
  • reaction solution was quenched by adding saturated aqueous ammonium chloride solution (20 mL), extracted with ethyl acetate (40 mL ⁇ 3), washed with saturated brine (20 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product.
  • Dissolve compound 14d (350 mg, 1.40 mmol) in dichloromethane (5 mL), add N,N-diisopropylethylamine (541.33 mg, 4.19 mmol, 729.56 ⁇ L), cool to 0°C, and add trifluoromethanesulfonate acid anhydride (787.85 mg, 2.79 mmol, 460.73 ⁇ L), and then reacted at 20°C for 0.5 hr. Add 5 mL of saturated ammonium chloride, extract with ethyl acetate (3 mL ⁇ 3), separate the liquids and combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product.
  • reaction solution was slowly poured into 120 mL of saturated ammonium chloride solution, and then extracted with ethyl acetate (50 mL Concentrate under reduced pressure to obtain crude product.

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Abstract

Disclosed are a series of aminopyrimidine compounds and use thereof, and specifically disclosed are a compound represented by formula (XI), a stereoisomer thereof and a pharmaceutically acceptable salt thereof, and use thereof.

Description

氨基嘧啶类化合物及其应用Aminopyrimidine compounds and their applications
本申请主张如下优先权：This application claims the following priority rights:
CN202210673201.2，申请日：2022年06月14日；CN202210673201.2, application date: June 14, 2022;
CN202211242420.1，申请日：2022年10月11日；CN202211242420.1, application date: October 11, 2022;
CN202211414875.7，申请日：2022年11月11日；CN202211414875.7, application date: November 11, 2022;
CN202211478036.1，申请日：2022年11月23日；CN202211478036.1, application date: November 23, 2022;
CN2023100420083，申请日：2023年01月12日；CN2023100420083, application date: January 12, 2023;
CN2023102778971，申请日：2023年03月21日。CN2023102778971, application date: March 21, 2023.
技术领域Technical field
本发明涉及一系列氨基嘧啶类化合物及其应用，具体涉及了式(XI)所示化合物、其立体异构体及其药学上可接受的盐及其应用。The present invention relates to a series of aminopyrimidine compounds and their applications, specifically to the compounds represented by formula (XI), their stereoisomers and their pharmaceutically acceptable salts and their applications.
背景技术Background technique
EGFR(Epidermal Growth Factor Receptor，简称为EGFR、ErbB-1或HER1)是表皮生长因子受体(HER)家族成员之一。EGFR广泛分布于哺乳动物上皮细胞、成纤维细胞、胶质细胞、角质细胞等细胞表面，EGFR信号通路对细胞的生长、增殖和分化等生理过程发挥重要作用。许多肿瘤中有突变型EGFR存在，现已发现许多种EGFR突变型。EGFR突变中的超过80％为19Del及L858R类型的突变，其余的属于罕见突变，主要包括18外显子上的G719X、E709X、Del18，19外显子上的Ins19，20外显子上的Ins20、S768I，21外显子上的L861Q。针对EGFR等基因驱动突变的患者，靶向治疗显示了其卓越的疗效。EGFR (Epidermal Growth Factor Receptor, referred to as EGFR, ErbB-1 or HER1) is a member of the epidermal growth factor receptor (HER) family. EGFR is widely distributed on the surface of mammalian epithelial cells, fibroblasts, glial cells, keratinocytes and other cells. The EGFR signaling pathway plays an important role in physiological processes such as cell growth, proliferation and differentiation. Mutated EGFR exists in many tumors, and many EGFR mutant types have been discovered. More than 80% of EGFR mutations are 19Del and L858R mutations, and the rest are rare mutations, mainly including G719X, E709X, and Del18 in exon 18, Ins19 in exon 19, and Ins20 in exon 20. , S768I, L861Q on exon 21. Targeted therapy has shown excellent efficacy for patients with driver mutations in genes such as EGFR.
当前EGFR靶向小分子酪氨酸激酶抑制剂(TKIs)有三代。第一代EGFR-TKIs，如吉非替尼，厄洛替尼，其采用与ATP竞争性非共价结合，对于在19号外显子的缺失突变和21号外显子的点突变的EGFR进行抑制。但一代EGFR-TKIs除了会竞争性抑制突变的EGFR蛋白，也能抑制正常细胞的EGFR功能，导致了皮疹，腹泻和无食欲这些副作用的产生。第二代EGFR-TKIs，如阿法替尼、达克替尼，其采用共价键结合的方式与ATP竞争性不可逆结合，抑制作用强度更大，无进展生存周期mPFS有所改善。缺点是由于不可逆的抑制，副作用更强。通常在服药一或二代EGFR抑制剂后第9.2到14.7个月时，EGFR-TKIs结合的氨基酸片段发生了T790M突变导致耐药，占比约有50-70％。There are currently three generations of EGFR-targeting small molecule tyrosine kinase inhibitors (TKIs). The first-generation EGFR-TKIs, such as gefitinib and erlotinib, use competitive non-covalent binding to ATP to inhibit EGFR with deletion mutations in exon 19 and point mutations in exon 21. . However, in addition to competitively inhibiting mutated EGFR proteins, first-generation EGFR-TKIs can also inhibit the EGFR function of normal cells, leading to side effects such as rash, diarrhea, and loss of appetite. Second-generation EGFR-TKIs, such as afatinib and dacomitinib, competitively and irreversibly bind to ATP through covalent bonding. The inhibitory effect is stronger and the progression-free survival period mPFS is improved. The disadvantage is that the side effects are stronger due to irreversible inhibition. Usually between 9.2 and 14.7 months after taking first- or second-generation EGFR inhibitors, the T790M mutation occurs in the amino acid fragment bound to EGFR-TKIs, leading to drug resistance, accounting for about 50-70%.
三代EGFR-TKIs阿斯利康的AZD9291(奥希替尼)成了首个针对EGFRT790M突变的抑制剂，靶向非小细胞肺癌的EGFR基因突变(包括Exon18,19,21突变)。用药10个月后，会带来外显子20C797突变，造成三代药物AZD9291耐药，对EGFR del19/L858R T790M C797S三突变疗效不够。因此开发对EGFR del19/L858R T790M C797S三突变作用，同时兼有对野生型EGFR选择性药物具有巨大的临床价值和市场价值。本发明化合物对EGFR del19/L858R T790M C797S三突变具有优异的抑制活性，同时对野生型EGFT良好的选择性，有望用于EGFR异常的疾病患者。The third-generation EGFR-TKIs AstraZeneca's AZD9291 (osimertinib) has become the first inhibitor to target the EGFRT790M mutation, targeting EGFR gene mutations in non-small cell lung cancer (including Exon18, 19, and 21 mutations). After 10 months of medication, the exon 20C797 mutation will occur, resulting in resistance to the third-generation drug AZD9291, and insufficient efficacy against the EGFR del19/L858R T790M C797S triple mutation. Therefore, the development of drugs that are selective for EGFR del19/L858R T790M C797S triple mutations while also targeting wild-type EGFR has huge clinical and market value. The compound of the present invention has excellent inhibitory activity against the EGFR del19/L858R T790M C797S triple mutation, and at the same time has good selectivity for wild-type EGFT, and is expected to be used in patients with EGFR abnormal diseases.
发明内容Contents of the invention
本发明提供了式(XI)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention provides the compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
其中，in,
T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
T4选自C和N；T 4 is selected from C and N;
T5选自CH和N；T 5 is selected from CH and N;
环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
环B选自吡唑基、哌啶基、四氢吡啶基、二氢吡喃基、环己基、环己烯基和7-10元杂环烷基；Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
R1选自-C(Ra)2S(＝O)2C(Rb)3、 R 1 is selected from -C(R a ) 2 S(=O) 2 C(R b ) 3 ,
R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
R3选自H、卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代；R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
R4选自F、OH、C1-3烷基、C1-3烷氧基、和C1-3烷氨基，所述C1-3烷基、C1-3烷氧基和C1-3烷氨基分别独立地任选被1、2或3个卤素取代；R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
R6和R7分别独立地选自H、卤素和C(Ra)3；R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
R8选自H和卤素；R 8 is selected from H and halogen;
T3选自C(Rc)2、NRc和O；T 3 is selected from C(R c ) 2 , NR c and O;
m选自0、1、2和3；m is selected from 0, 1, 2 and 3;
n选自1、2和3；n is selected from 1, 2 and 3;
r选自1、2和3；r is selected from 1, 2 and 3;
s选自1和2；s is selected from 1 and 2;
t选自1和2；t is selected from 1 and 2;
各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
Rc选自H和C1-3烷基；R c is selected from H and C 1-3 alkyl;
条件是， requirement is,
1)当R1选自-C(Ra)2S(＝O)2C(Rb)3时，环A选自2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；1) When R 1 is selected from -C(R a ) 2 S(=O) 2 C(R b ) 3 , ring A is selected from 2,3-dihydrobenzofuryl, phthalyl, 2,3 -Dihydropyridofuranyl and isoindolinyl;
2)当R1选自时，环B不选自吡唑基。2) When R 1 is selected from When , Ring B is not selected from pyrazolyl.
本发明还提供了式(XI)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention also provides the compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
其中，in,
T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
T4选自C和N；T 4 is selected from C and N;
T5选自CH和N；T 5 is selected from CH and N;
环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
环B选自吡唑基、哌啶基、四氢吡啶基、二氢吡喃基、环己基、环己烯基和7-10元杂环烷基；Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
R1选自C1-3烷基、-C(C(Ra)3)2(OH)、-C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2P(＝O)(C(Rb)3)2、S(＝O)2C(Rb)3、所述C1-3烷基任选被1、2、3、4或5个卤素取代；R 1 is selected from C 1-3 alkyl, -C(C(R a ) 3 ) 2 (OH), -C(R a ) 2 S(=O) 2 C(R b ) 3 , -C(R a ) 2 C(R a ) 2 S(＝O) 2 C(R b ) 3 , -C(R a ) 2 P(＝O)(C(R b ) 3 ) 2 , S(＝O) 2 C(R b ) 3 , The C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
R3选自H、卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代；R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
R4选自F、OH、C1-3烷基、C1-3烷氧基、和C1-3烷氨基，所述C1-3烷基、C1-3烷氧基和C1-3烷氨基分别独立地任选被1、2或3个卤素取代；R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
R6和R7分别独立地选自H、卤素和C(Ra)3； R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
R8选自H和卤素；R 8 is selected from H and halogen;
T3选自CRc、NRc和O；T 3 is selected from CR c , NR c and O;
m选自0、1、2和3；m is selected from 0, 1, 2 and 3;
n选自1、2和3；n is selected from 1, 2 and 3;
r选自1、2和3；r is selected from 1, 2 and 3;
s选自1和2；s is selected from 1 and 2;
t选自1和2；t is selected from 1 and 2;
各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
Rc选自H和C1-3烷基；R c is selected from H and C 1-3 alkyl;
本发明还提供了式(XI)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention also provides the compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
其中，in,
T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
T4选自C和N；T 4 is selected from C and N;
T5选自CH和N；T 5 is selected from CH and N;
环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
环B选自吡唑基、哌啶基、四氢吡啶基、二氢吡喃基、环己基、环己烯基和7-10元杂环烷基；Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
R1选自-C(Ra)2S(＝O)2C(Rb)3、 R 1 is selected from -C(R a ) 2 S(=O) 2 C(R b ) 3 ,
R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
R3选自H、卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代； R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
R4选自F、OH、C1-3烷基、C1-3烷氧基、和C1-3烷氨基，所述C1-3烷基、C1-3烷氧基和C1-3烷氨基分别独立地任选被1、2或3个卤素取代；R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
R6和R7分别独立地选自H、卤素和C(Ra)3；R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
R8选自H和卤素；R 8 is selected from H and halogen;
T3选自CRc、NRc和O；T 3 is selected from CR c , NR c and O;
m选自0、1、2和3；m is selected from 0, 1, 2 and 3;
n选自1、2和3；n is selected from 1, 2 and 3;
r选自1、2和3；r is selected from 1, 2 and 3;
s选自1和2；s is selected from 1 and 2;
t选自1和2；t is selected from 1 and 2;
各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
Rc选自H和C1-3烷基；R c is selected from H and C 1-3 alkyl;
条件是，requirement is,
1)当R1选自-C(Ra)2S(＝O)2C(Rb)3时，环A选自2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；1) When R 1 is selected from -C(R a ) 2 S(=O) 2 C(R b ) 3 , ring A is selected from 2,3-dihydrobenzofuryl, phthalyl, 2,3 -Dihydropyridofuranyl and isoindolinyl;
2)当R1选自时，环B不选自吡唑基。2) When R 1 is selected from When , Ring B is not selected from pyrazolyl.
本发明还提供了式(V)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention also provides the compound represented by formula (V), its stereoisomer or its pharmaceutically acceptable salt,
其中，in,
T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
T4选自C和N；T 4 is selected from C and N;
T5选自CH和N； T 5 is selected from CH and N;
环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
环B选自吡唑基、哌啶基、四氢吡啶基、二氢吡喃基、环己基、环己烯基和7-10元杂环烷基；Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
R1选自C1-3烷基、-C(C(Ra)3)2(OH)、-C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2P(＝O)(C(Rb)3)2、S(＝O)2C(Rb)3、所述C1-3烷基任选被1、2、3、4或5个卤素取代；R 1 is selected from C 1-3 alkyl, -C(C(R a ) 3 ) 2 (OH), -C(R a ) 2 S(=O) 2 C(R b ) 3 , -C(R a ) 2 C(R a ) 2 S(＝O) 2 C(R b ) 3 , -C(R a ) 2 P(＝O)(C(R b ) 3 ) 2 , S(＝O) 2 C(R b ) 3 , The C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
T3选自NRc和O；T 3 is selected from NR c and O;
R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
R3选自H、卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代；R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
R4选自F、OH、CH3、OCH3、CH2CH3和N(CH3)2；R 4 is selected from F, OH, CH 3 , OCH 3 , CH 2 CH 3 and N(CH 3 ) 2 ;
R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
R6和R7分别独立地选自H、卤素和C(Ra)3；R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
m选自0、1、2和3；m is selected from 0, 1, 2 and 3;
n选自1、2和3；n is selected from 1, 2 and 3;
r选自1、2和3；r is selected from 1, 2 and 3;
s选自1和2；s is selected from 1 and 2;
t选自1和2；t is selected from 1 and 2;
各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
Rc选自H和C1-3烷基；R c is selected from H and C 1-3 alkyl;
条件是，requirement is,
当R1选自-CH2S(＝O)2C(Rb)3，Rb选自H，T1选自N，T2选自N，T4选自C，且结构单元选自 其中T7和T8分别独立地选自N和CR，R选自H，卤素和CH3时，When R 1 is selected from -CH 2 S(=O) 2 C(R b ) 3 , R b is selected from H, T 1 is selected from N, T 2 is selected from N, T 4 is selected from C, and the structural unit Selected from Where T 7 and T 8 are independently selected from N and CR, and R is selected from H, halogen and CH 3 ,
1)环B选自吡唑基四氢吡啶基、二氢吡喃基、环己基和环己烯基，或者；1) Ring B is selected from pyrazolyltetrahydropyridyl, dihydropyranyl, cyclohexyl and cyclohexenyl, or;
2)当R2选自H，R6选自H时，R7不为H且不为CH3，或者；2) When R 2 is selected from H and R 6 is selected from H, R 7 is not H and not CH 3 , or;
3)当R2选自H，R7选自H时，R6不为H且不为CH3。3) When R 2 is selected from H and R 7 is selected from H, R 6 is not H and is not CH 3 .
本发明提供了式(V)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention provides the compound represented by formula (V), its stereoisomer or its pharmaceutically acceptable salt,
其中，in,
T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
T4选自C和N；T 4 is selected from C and N;
T5选自CH和N；T 5 is selected from CH and N;
环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl, and isoindoline;
环B选自吡唑基和哌啶基；Ring B is selected from pyrazolyl and piperidinyl;
R1选自C1-3烷基、-C(C(Ra)3)2(OH)、-C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2P(＝O)(C(Rb)3)2、S(＝O)2C(Rb)3、所述C1-3烷基任选被1、2、3、4或5个卤素取代；R 1 is selected from C 1-3 alkyl, -C(C(R a ) 3 ) 2 (OH), -C(R a ) 2 S(=O) 2 C(R b ) 3 , -C(R a ) 2 C(R a ) 2 S(＝O) 2 C(R b ) 3 , -C(R a ) 2 P(＝O)(C(R b ) 3 ) 2 , S(＝O) 2 C(R b ) 3 , The C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
T3选自NRc和O；T 3 is selected from NR c and O;
R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
R3选自H、卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代；R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
R4选自F、OH、CH3、OCH3和 R 4 is selected from F, OH, CH 3 , OCH 3 and
R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
R6和R7分别独立地选自H、卤素和C(Ra)3；R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
m选自1、2和3；m is selected from 1, 2 and 3;
n选自1、2和3；n is selected from 1, 2 and 3;
r选自1、2和3；r is selected from 1, 2 and 3;
s选自1和2；s is selected from 1 and 2;
t选自1和2；t is selected from 1 and 2;
各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
Rc选自H和C1-3烷基；R c is selected from H and C 1-3 alkyl;
条件是，requirement is,
当R1选自-CH2S(＝O)2C(Rb)3，T1选自N，T2选自N，T4选自C，且结构单元选自其中T7和T8分别独立地选自N和CR，R选自H，卤素和CH3时，When R 1 is selected from -CH 2 S(=O) 2 C(R b ) 3 , T 1 is selected from N, T 2 is selected from N, T 4 is selected from C, and the structural unit Selected from Where T 7 and T 8 are independently selected from N and CR, and R is selected from H, halogen and CH 3 ,
1)环B选自吡唑基，或者；1) Ring B is selected from pyrazolyl, or;
2)当R2选自H，R6选自H时，R7不为H且不为CH3，或者；2) When R 2 is selected from H and R 6 is selected from H, R 7 is not H and not CH 3 , or;
3)当R2选自H，R7选自H时，R6不为H且不为CH3。3) When R 2 is selected from H and R 7 is selected from H, R 6 is not H and is not CH 3 .
本发明还提供了式(V)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention also provides the compound represented by formula (V), its stereoisomer or its pharmaceutically acceptable salt,
其中，in,
T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
T4选自C和N；T 4 is selected from C and N;
T5选自CH和N；T 5 is selected from CH and N;
环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基和2,3-二氢吡啶并呋喃基；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- Dihydrobenzofuryl, phthalyl and 2,3-dihydropyridofuranyl;
环B选自吡唑基和哌啶基；Ring B is selected from pyrazolyl and piperidinyl;
R1选自C1-3烷基、-C(C(Ra)3)2(OH)、-C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2C(Ra)2S(＝O)2C(Rb)3、-C(Ra)2P(＝O)(C(Rb)3)2、 S(＝O)2C(Rb)3、所述C1-3烷基任选被1、2、3、4或5个卤素取代；R 1 is selected from C 1-3 alkyl, -C(C(R a ) 3 ) 2 (OH), -C(R a ) 2 S(=O) 2 C(R b ) 3 , -C(R a ) 2 C(R a ) 2 S(＝O) 2 C(R b ) 3 , -C(R a ) 2 P(＝O)(C(R b ) 3 ) 2 , S(=O) 2 C(R b ) 3 , The C 1-3 alkyl group is optionally substituted by 1, 2, 3, 4 or 5 halogens;
T3选自NRc和O；T 3 is selected from NR c and O;
R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
R3选自卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代；R 3 is selected from halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optional Substituted by 1, 2 or 3 halogens;
R4选自F、OH、CH3、OCH3和 R 4 is selected from F, OH, CH 3 , OCH 3 and
R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
R6和R7分别独立地选自H、卤素和C(Ra)3；R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
m选自1、2和3；m is selected from 1, 2 and 3;
n选自1、2和3；n is selected from 1, 2 and 3;
s选自1和2；s is selected from 1 and 2;
t选自1和2；t is selected from 1 and 2;
各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
Rc选自H和CH3；R c is selected from H and CH 3 ;
条件是，requirement is,
当R1选自-CH2S(＝O)2C(Rb)3，环A选自苯基和吡啶基时，When R 1 is selected from -CH 2 S(=O) 2 C(R b ) 3 and ring A is selected from phenyl and pyridyl,
1)环B选自吡唑基，或者1) Ring B is selected from pyrazolyl, or
2)R2、R6、R7和Ra不同时选自H，或者2) R 2 , R 6 , R 7 and R a are not selected from H at the same time, or
3)T2选自CH。3) T 2 is selected from CH.
在本发明的一些方案中，上述T3选自CH2、NH、NCH3、NCH2CH3、NCH(CH3)2和O，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned T 3 is selected from CH 2 , NH, NCH 3 , NCH 2 CH 3 , NCH(CH 3 ) 2 and O, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R1选自CH3、-C(CH3)2(OH)、-CH2S(＝O)2CH3、-CH2CH2S(＝O)2CH3、-CHFS(＝O)2CH3、-CF2S(＝O)2CH3、-CH2P(＝O)(CH3)2、-S(＝O)2CH3、 其他变量如本发明所定义。In some aspects of the invention, the above R 1 is selected from CH 3 , -C(CH 3 ) 2 (OH), -CH 2 S(=O) 2 CH 3 , -CH 2 CH 2 S(=O) 2 CH 3 , -CHFS(=O) 2 CH 3 , -CF 2 S(=O) 2 CH 3 , -CH 2 P(=O)(CH 3 ) 2 , -S(=O) 2 CH 3 , Other variables are as defined in the present invention.
在本发明的一些方案中，上述R1选自-CH2S(＝O)2CH3、-CHFS(＝O)2CH3、-CF2S(＝O)2CH3、 其他变量如本发明所定义。In some aspects of the invention, the above-mentioned R 1 is selected from -CH 2 S(=O) 2 CH 3 , -CHFS(=O) 2 CH 3 , -CF 2 S(=O) 2 CH 3 , Other variables are as defined in the present invention.
在本发明的一些方案中，上述R1选自CH3、-C(CH3)2(OH)、-CH2S(＝O)2CH3、-CH2CH2S(＝O)2CH3、-CHFS(＝O)2CH3、-CF2S(＝O)2CH3、-CH2P(＝O)(CH3)2、-S(＝O)2CH3、 其他变量如本发明所定义。In some aspects of the invention, the above R 1 is selected from CH 3 , -C(CH 3 ) 2 (OH), -CH 2 S(=O) 2 CH 3 , -CH 2 CH 2 S(=O) 2 CH 3 , -CHFS(=O) 2 CH 3 , -CF 2 S(=O) 2 CH 3 , -CH 2 P(=O)(CH 3 ) 2 , -S(=O) 2 CH 3 , Other variables are as defined in the present invention.
在本发明的一些方案中，上述R2选自H、F和OH，或者不存在，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned R 2 is selected from H, F and OH, or does not exist, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R5选自＝CHS(＝O)2CH3、 其他变量如本发明所定义。In some aspects of the invention, the above-mentioned R 5 is selected from =CHS(=O) 2 CH 3 , Other variables are as defined in the present invention.
在本发明的一些方案中，上述R5选自＝CHS(＝O)2CH3、其他变量如本发明所定义。In some aspects of the invention, the above-mentioned R 5 is selected from =CHS(=O) 2 CH 3 , Other variables are as defined in the present invention.
在本发明的一些方案中，上述R6和R7分别独立地选自H和CH3，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned R 6 and R 7 are independently selected from H and CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R6选自H和CH3，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned R 6 is selected from H and CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R7选自H和CH3，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned R 7 is selected from H and CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R8选自H和F，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 8 is selected from H and F, and other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自其他变量如本发明所定义。 In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。 In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述R3选自F、CH3、CH2CH3、CH(CH3)2、OCH3和环丙基，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned R 3 is selected from F, CH 3 , CH 2 CH 3 , CH(CH 3 ) 2 , OCH 3 and cyclopropyl, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R3选自F、CH3、CH(CH3)2、OCH3和环丙基，其他变量如本发明所定义。In some aspects of the present invention, the above-mentioned R 3 is selected from F, CH 3 , CH(CH 3 ) 2 , OCH 3 and cyclopropyl, and other variables are as defined in the present invention.
在本发明的一些方案中，上述环A选自2,3-二氢苯并呋喃基、苯基和吡啶基，其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned Ring A is selected from 2,3-dihydrobenzofuranyl, phenyl and pyridyl, and other variables are as defined in the present invention.
在本发明的一些方案中，上述环A选自苯基和吡啶基，其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A is selected from phenyl and pyridyl, and other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。 In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述环B选自 其他变量如本发明所定义。In some aspects of the invention, the above-mentioned ring B is selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述环B选自 其他变量如本发明所定义。In some aspects of the invention, the above-mentioned ring B is selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述环B选自哌啶基、四氢吡啶基、二氢吡喃基、环己烯基和7-10元杂环烷基，其他变量如本发明所定义。In some aspects of the invention, the above-mentioned ring B is selected from piperidyl, tetrahydropyridyl, dihydropyranyl, cyclohexenyl and 7-10 membered heterocycloalkyl, and other variables are as defined in the invention.
在本发明的一些方案中，上述R4选自F、OH、CH3、OCH3、CH2CH3、N(CH3)2和其他变量如本发明所定义。In some aspects of the invention, the above-mentioned R 4 is selected from F, OH, CH 3 , OCH 3 , CH 2 CH 3 , N(CH 3 ) 2 and Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自其他变量如本 发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as follows defined by invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。 In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some solutions of the present invention, the above structural units Selected from Other variables are as defined in the present invention.
在本发明的一些方案中，上述化合物、其立体异构体或其药学上可接受的盐，其选自，
In some aspects of the present invention, the above compounds, their stereoisomers or their pharmaceutically acceptable salts are selected from,
其中，R1选自 Among them, R 1 is selected from
环A、环B、R2、R3、R4、R6、R7、R8、T1、T2、T3、T4、T5、m、n和r如本发明所定义。Ring A, Ring B, R2 , R3 , R4, R6 , R7 , R8 , T1 , T2 , T3 , T4 , T5 , m, n and r are as defined herein.
在本发明的一些方案中，上述化合物、其立体异构体或其药学上可接受的盐，其选自，
In some aspects of the present invention, the above compounds, their stereoisomers or their pharmaceutically acceptable salts are selected from,
其中， in,
R1选自环A、R2、R3、R4、R6、R7、T3、m、n和r如本发明所定义。R 1 is selected from Rings A, R2 , R3 , R4 , R6 , R7 , T3 , m, n and r are as defined herein.
本发明还有一些方案由上述变量任意组合而来。There are also some solutions of the present invention derived from any combination of the above variables.
本发明提供了下列化合物、其立体异构体或其药学上可接受的盐，











The present invention provides the following compounds, their stereoisomers or their pharmaceutically acceptable salts,











在本发明的一些方案中，所述化合物、其立体异构体或其药学上可接受的盐，其化合物选自，





































In some aspects of the invention, the compound, its stereoisomer or its pharmaceutically acceptable salt, the compound thereof is selected from,





































在本发明的一些方案中，所述化合物、其立体异构体或其药学上可接受的盐，其化合物选自，



In some aspects of the invention, the compound, its stereoisomer or its pharmaceutically acceptable salt, the compound thereof is selected from,



本发明还提供了一种药物组合物，包括治疗有效量的如本发明所定义的化合物、其立体异构体或其药学上可接受的盐作为活性成分以及药学上可接受的载体、稀释剂或赋形剂。The present invention also provides a pharmaceutical composition, including a therapeutically effective amount of a compound as defined in the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier and diluent. or excipients.
本发明还提供了上述化合物、其立体异构体或其药学上可接受的盐或者上述组合物在制备治疗实体瘤药物中的应用。The present invention also provides the use of the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt, or the above-mentioned composition in the preparation of drugs for treating solid tumors.
在本发明的一些方案中，所述实体瘤指非小细胞肺癌等实体瘤。In some aspects of the present invention, the solid tumor refers to solid tumors such as non-small cell lung cancer.
本发明还提供了上述化合物的下列合成方法：The present invention also provides the following synthesis methods of the above compounds:
合成方法1
Synthesis method 1
合成方法2
Synthesis method 2
本发明还提供了上述化合物的下列测试方法： The present invention also provides the following test methods for the above compounds:
测试方法1：化合物对EGFR(T790M/C797S/L858R)的激酶抑制活性Test method 1: Kinase inhibitory activity of compounds against EGFR (T790M/C797S/L858R)
在反应缓冲液(20mM羟乙基哌嗪乙硫磺酸(Hepes)(pH 7.5),10mM氯化镁(MgCl2),1mM乙二醇双氨乙基醚四乙酸(EGTA),0.02％聚氧乙烯十二烷醚(Brij35),0.02mg/mL BSA,0.1mM钒酸钠(Na3VO4),2mM二硫苏糖醇(DTT),1％DMSO)中依次加入一定浓度的底物、辅酶因子、激酶和测试化合物(10个剂量，3倍连续稀释液，2％DMSO最终浓度)并混匀，将混合物在室温下孵育20分钟，向反应混合液中加入一定浓度的33P-ATP开始反应，随后室温孵育120分钟。最后通过过滤-结合的方法来检测反应物的放射性。最终的激酶活性以测试样品中剩余的激酶活性占DMSO对照组的激酶活性的比例来表示。通过GraphPad软件拟合量效关系曲线并计算IC50。In reaction buffer (20mM hydroxyethyl piperazine ethyl sulfonic acid (Hepes) (pH 7.5), 10mM magnesium chloride (MgCl 2 ), 1mM ethylene glycol bisaminoethyl ether tetraacetic acid (EGTA), 0.02% polyoxyethylene ten Dialkyl ether (Brij35), 0.02mg/mL BSA, 0.1mM sodium vanadate (Na 3 VO 4 ), 2mM dithiothreitol (DTT), 1% DMSO) and then add a certain concentration of substrate and coenzyme factor , kinase and test compound (10 doses, 3-fold serial dilution, 2% DMSO final concentration) and mix well, incubate the mixture at room temperature for 20 minutes, add a certain concentration of 33 P-ATP to the reaction mixture to start the reaction , followed by incubation at room temperature for 120 minutes. Finally, the radioactivity of the reactants is detected by filtration-binding method. The final kinase activity is expressed as the ratio of the remaining kinase activity in the test sample to the kinase activity in the DMSO control group. The dose-response relationship curve was fitted and IC 50 was calculated using GraphPad software.
测试方法2：化合物对Ba/F3-FL-EGFR(T790M/C797S/L858R)细胞增殖的抑制活性Test method 2: Inhibitory activity of compounds on Ba/F3-FL-EGFR (T790M/C797S/L858R) cell proliferation
三磷酸腺苷(Adenosine Tri-Phosphate,ATP)是自然界中各种生命活动中共用的能量载体，是能量储存和转移的最小单位。CellTiter-GloTM活细胞检测试剂盒采用萤光素酶作检测物，发光过程中萤光素酶需要ATP的参与。向细胞培养基中加入CellTiter-GloTM试剂，测量发光值，光信号和体系中ATP量成正比，而ATP又和活细胞数正相关。因此通过使用CellTiter-Glo试剂盒检测ATP含量，可以检测出细胞的增殖情况。本测试中，细胞系为Ba/F3-FL-EGFR(T790M/C797S/L858R)稳转细胞株。Adenosine Tri-Phosphate (ATP) is a common energy carrier in various life activities in nature and is the smallest unit of energy storage and transfer. CellTiter-Glo TM live cell detection kit uses luciferase as the detection substance, and luciferase requires the participation of ATP during the luminescence process. Add CellTiter-Glo TM reagent to the cell culture medium and measure the luminescence value. The light signal is directly proportional to the amount of ATP in the system, and ATP is positively correlated with the number of viable cells. Therefore, by using the CellTiter-Glo kit to detect ATP content, cell proliferation can be detected. In this test, the cell line was Ba/F3-FL-EGFR (T790M/C797S/L858R) stably transfected cell line.
IC50测定过程：IC 50 determination process:
1)细胞培养1) Cell culture
将细胞系在培养条件37℃，5％CO2的培养箱中进行培养。定期传代，取处于对数生长期的细胞用于铺板。The cell lines were cultured in an incubator with culture conditions of 37°C and 5% CO2 . Passage regularly and use cells in the logarithmic growth phase for plating.
2)化合物存储板制备2) Compound storage plate preparation
a)用DMSO将待测化合物配置成10mM溶液，再用DMSO将待测化合物稀释至0.3或1mM。a) Use DMSO to prepare the compound to be tested into a 10mM solution, and then use DMSO to dilute the compound to be tested to 0.3 or 1mM.
b)制备1000×化合物存储板(管)：用DMSO从最高浓度3倍梯度稀释至最低浓度,9个浓度。b) Prepare 1000× compound storage plate (tube): use DMSO to dilute 3-fold gradient from the highest concentration to the lowest concentration, 9 concentrations.
c)20×化合物工作液的配制:在平底的96孔透明药板中加入49μL细胞培养液，从1000×化合物存储板中吸取1μL化合物加入96孔透明药板的细胞培养液中。在溶媒对照中加入1μL DMSO。加入化合物或DMSO后用排枪吹打混匀。c) Preparation of 20× compound working solution: Add 49 μL of cell culture medium to a flat-bottomed 96-well transparent medicine plate, pipet 1 μL of compound from the 1000× compound storage plate and add it to the cell culture medium of the 96-well transparent medicine plate. Add 1 μL DMSO to the vehicle control. Add the compound or DMSO and mix with pipette.
3)细胞铺板与给药3) Cell plating and drug administration
a)用台盼兰进行细胞染色并计数活细胞，要求细胞活率90％以上。a) Use trypan blue to stain cells and count viable cells. The cell viability rate is required to be above 90%.
b)在化合物检测细胞板中每孔加入95μL细胞悬液(2000cells/well)，在Min对照孔中加入不含细胞(含0.1％DMSO)的培养液。b) Add 95 μL of cell suspension (2000 cells/well) to each well of the compound detection cell plate, and add culture medium without cells (containing 0.1% DMSO) to the Min control well.
c)化合物检测细胞板加药：取5μL的20×化合物工作液按表1所示加入到细胞培养板中。在Max对照中加入5μL DMSO-细胞培养液混合液。DMSO终浓度为0.1％。c) Compound detection cell plate addition: Take 5 μL of 20× compound working solution and add it to the cell culture plate as shown in Table 1. Add 5 μL of DMSO-cell culture medium mixture to the Max control. The final DMSO concentration is 0.1%.
d)将培养板在37℃,5％CO2培养箱中培养72小时。d) Incubate the culture plate in a 37°C, 5% CO 2 incubator for 72 hours.
4)CellTiter-Glo发光法细胞活性检测4)CellTiter-Glo luminescence method cell viability detection
以下步骤按照Promega CellTiter-Glo发光法细胞活性检测试剂盒(Promega-G7573)的说明书来进行。The following steps are carried out according to the instructions of Promega CellTiter-Glo Luminescence Cell Viability Detection Kit (Promega-G7573).
a)将CellTiter-Glo缓冲液融化并放置至室温。a) Melt CellTiter-Glo buffer and bring to room temperature.
b)将CellTiter-Glo底物放置至室温。b) Let CellTiter-Glo substrate come to room temperature.
c)在一瓶CellTiter-Glo底物中加入CellTiter-Glo缓冲液以溶解底物，从而配制CellTiter-Glo工作液。c) Add CellTiter-Glo buffer to a bottle of CellTiter-Glo substrate to dissolve the substrate to prepare CellTiter-Glo working solution.
d)缓慢涡旋震荡使充分溶解。d) Vortex slowly to fully dissolve.
e)取出细胞培养板放置10分钟使其平衡至室温。e) Take out the cell culture plate and let it equilibrate to room temperature for 10 minutes.
f)在每孔中加入50μL(等于每孔中细胞培养液一半体积)的CellTiter-Glo工作液。f) Add 50 μL (equal to half the volume of cell culture medium in each well) of CellTiter-Glo working solution into each well.
g)将培养板在轨道摇床上振摇2分钟以诱导细胞裂解。 g) Shake the plate on an orbital shaker for 2 minutes to induce cell lysis.
h)培养板在室温放置10分钟以稳定发光信号。h) Place the culture plate at room temperature for 10 minutes to stabilize the luminescence signal.
i)在PerkinElmer2105读板器上检测发光信号。i) At PerkinElmer The luminescence signal is detected on the 2105 plate reader.
5)数据处理5)Data processing
PerkinElmer2105读数得出对应的每孔荧光值RLU。PerkinElmer 2105 readings yield the corresponding fluorescence value RLU for each well.
细胞增殖抑制率(Inhibition Rate)数据采用下列公式来处理：Cell proliferation inhibition rate (Inhibition Rate) data is processed using the following formula:
Inhibition Rate(Inh％)＝100-(RLUDrug-RLUMin)/(RLUMax-RLUMin)*100％。Inhibition Rate(Inh%)=100-(RLUDrug-RLUMin)/(RLUMax-RLUMin)*100%.
在EXCEL中计算不同浓度化合物对应的抑制率，然后用GraphPad Prism软件作抑制率曲线图并计算IC50值。Calculate the inhibition rates corresponding to different concentrations of compounds in EXCEL, and then use GraphPad Prism software to draw the inhibition rate curve and calculate the IC 50 value.
相关定义Related definitions
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear in the absence of a specific definition, but should be understood in its ordinary meaning. Where a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue. , without undue toxicity, irritation, allergic reactions, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物接触的方式获得碱加成盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物接触的方式获得酸加成盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。The term "pharmaceutically acceptable salts" refers to salts of compounds of the present invention prepared from compounds having specific substituents found in the present invention and relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in pure solution or in a suitable inert solvent. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base or acid addition salts.
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。The pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid groups or bases. In general, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereoisomers isomer, the (D)-isomer, the (L)-isomer, as well as their racemic mixtures and other mixtures, such as enantiomeric or diastereomerically enriched mixtures, all of which belong to the present invention. within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
除非另有说明，术语“对映异构体”或者“旋光异构体”是指互为镜像关系的立体异构体。Unless otherwise stated, the terms "enantiomers" or "optical isomers" refer to stereoisomers that are mirror images of each other.
除非另有说明，术语“顺反异构体”或者“几何异构体”系由因双键或者成环碳原子单键不能自由旋转而引起。Unless otherwise stated, the term "cis-trans isomers" or "geometric isomers" refers to the inability of the double bonds or single bonds of the carbon atoms in the ring to rotate freely.
除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。Unless otherwise stated, the term "diastereomer" refers to stereoisomers whose molecules have two or more chiral centers and are in a non-mirror image relationship between the molecules.
除非另有说明，“(+)”表示右旋，“(-)”表示左旋，“(±)”表示外消旋。Unless otherwise stated, "(+)" means right-handed, "(-)" means left-handed, and "(±)" means racemic.
除非另有说明，用楔形实线键和楔形虚线键表示一个立体中心的绝对构型，用直形实线键和直形虚线键表示立体中心的相对构型，用波浪线表示楔形实线键或楔形虚线键或用波浪线表示直形实线键和直形虚线键如代 表如代表的混合物。Unless otherwise stated, use wedge-shaped solid line keys and wedge-shaped dotted keys Represents the absolute configuration of a three-dimensional center, using straight solid line keys and straight dotted keys Represent the relative configuration of the three-dimensional center with a wavy line Represents wedge-shaped solid line key or wedge-shaped dotted key or use tilde Represents a straight solid line key and straight dotted keys like generation surface like represent mixture.
除非另有说明，术语“互变异构体”或“互变异构体形式”是指在室温下，不同官能团异构体处于动态平衡，并能很快的相互转化。若互变异构体是可能的(如在溶液中)，则可以达到互变异构体的化学平衡。例如，质子互变异构体(proton tautomer)(也称质子转移互变异构体(prototropic tautomer))包括通过质子迁移来进行的互相转化，如酮-烯醇异构化和亚胺-烯胺异构化。价键异构体(valence tautomer)包括一些成键电子的重组来进行的相互转化。其中酮-烯醇互变异构化的具体实例是戊烷-2,4-二酮与4-羟基戊-3-烯-2-酮两个互变异构体之间的互变。Unless otherwise stated, the term "tautomer" or "tautomeric form" means that at room temperature, isomers with different functional groups are in dynamic equilibrium and can quickly convert into each other. If tautomers are possible (eg in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also called proton transfer tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enol isomerization. Amine isomerization. Valence tautomers include interconversions through the reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the tautomerization between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％，或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。Unless otherwise stated, the terms "enriched in an isomer," "enantiomerically enriched," "enriched in an enantiomer," or "enantiomerically enriched" refer to one of the isomers or enantiomers. The content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。Unless otherwise stated, the term "isomeric excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, then the isomer or enantiomeric excess (ee value) is 80% .
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。The optically active (R)- and (S)-isomers as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliaries, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with a suitable optically active acid or base, and then the salt is formed by conventional methods known in the art. Diastereomeric resolution is performed and the pure enantiomers are recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using chiral stationary phases, optionally combined with chemical derivatization methods (e.g., generation of amino groups from amines). formate).
本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚(3H)，碘-125(125I)或C-14(14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。The compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds can be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or C-14 ( 14 C). For another example, deuterated drugs can be replaced by heavy hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs can reduce side effects and increase drug stability. , enhance efficacy, extend drug biological half-life and other advantages. All variations in the isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
术语“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。The terms "optionally" or "optionally" mean that the subsequently described event or condition may, but need not, occur, and that the description includes instances in which the stated event or condition occurs and instances in which it does not. .
术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，取代基可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意味着两个氢原子被取代。氧取代不会发生在芳香基上。The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e. =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups.
术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。The term "optionally substituted" means that it may or may not be substituted. Unless otherwise specified, the type and number of substituents may be arbitrary on the basis of chemical achievability.
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。 When any variable (e.g., R) occurs more than once in the composition or structure of a compound, its definition in each instance is independent. Thus, for example, if a group is substituted by 0-2 R, then said group may optionally be substituted by up to two R's, with independent options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permitted only if such combinations result in stable compounds.
当一个连接基团的数量为0时，比如-(CRR)0-，表示该连接基团为单键。When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
当其中一个变量选自单键时，表示其连接的两个基团直接相连，比如A-L-Z中L代表单键时表示该结构实际上是A-Z。When one of the variables is selected from a single bond, it means that the two groups it is connected to are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.
除非另有规定，当某一基团具有一个或多个可连接位点时，该基团的任意一个或多个位点可以通过化学键与其他基团相连。当该化学键的连接方式是不定位的，且可连接位点存在H原子时，则连接化学键时，该位点的H原子的个数会随所连接化学键的个数而对应减少变成相应价数的基团。所述位点与其他基团连接的化学键可以用直形实线键直形虚线键或波浪线表示。例如-OCH3中的直形实线键表示通过该基团中的氧原子与其他基团相连；中的直形虚线键表示通过该基团中的氮原子的两端与其他基团相连；中的波浪线表示通过该苯基基团中的1和2位碳原子与其他基团相连；表示该哌啶基上的任意可连接位点可以通过1个化学键与其他基团相连，至少包括 这4种连接方式，即使-N-上画出了H原子，但是仍包括这种连接方式的基团，只是在连接1个化学键时，该位点的H会对应减少1个变成相应的一价哌啶基；表示R可以任意连接在双键的两端，即表示 Unless otherwise specified, when a certain group has one or more attachable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the connection mode of the chemical bond is non-positioned and there are H atoms at the connectable site, when the chemical bond is connected, the number of H atoms at the site will be reduced correspondingly with the number of connected chemical bonds and become the corresponding valence. group. The chemical bond connecting the site to other groups can be a straight solid line bond straight dashed key or wavy lines express. For example, the straight solid line bond in -OCH 3 means that it is connected to other groups through the oxygen atom in the group; The straight dotted bond in means that it is connected to other groups through both ends of the nitrogen atoms in the group; The wavy lines in indicate that the phenyl group is connected to other groups through the 1 and 2 carbon atoms in the phenyl group; Indicates that any connectable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if H atoms are drawn on -N-, still includes For groups with this connection method, when connecting a chemical bond, the H at that position will be reduced by one and become the corresponding monovalent piperidinyl group; It means that R can be connected at both ends of the double bond arbitrarily, which means
除非另有规定，术语“C1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C1-3烷基包括C1-2和C2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C1-3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。Unless otherwise specified, the term "C 1-3 alkyl" is used to mean a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) . Examples of C 1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.
除非另有规定，术语“卤代素”或“卤素”本身或作为另一取代基的一部分表示氟、氯、溴或碘原子。Unless otherwise specified, the term "halogen" or "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
除非另有规定，术语“C1-3烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C1-3烷氧基包括C1-2、C2-3、C3和C2烷氧基等。C1-3烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)等。Unless otherwise specified, the term "C 1-3 alkoxy" means those alkyl groups containing 1 to 3 carbon atoms that are attached to the remainder of the molecule through an oxygen atom. The C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy groups, etc. Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
除非另有规定，术语“7-10元杂环烷基”本身或者与其他术语联合分别表示由7至10个环原子组成的饱和环状基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子，其中氮原子任选地被季铵化，碳、氮和硫杂原子可任选被氧化(即C(＝O)、NO和S(O)p，p是1或2)。其包括单环、双环和三环体系，其中双环和三环体系包括螺环、并环和桥环。此外，就该“7-10元杂环烷基”而言，杂原子可以占据杂环烷基与分子其余部分的连接位置。所述7-10元杂环烷基包括7-10元、7-9元、7-8元、8-10元、8-9元、8元、9元和10元杂环烷基等。7-10元杂环烷基的实例包括但不限于 等。Unless otherwise specified, the term "7-10 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 7 to 10 ring atoms, with 1, 2, 3 or 4 ring atoms. are heteroatoms independently selected from O, S and N, and the remainder are carbon atoms, in which the nitrogen atoms are optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms can be optionally oxidized (i.e. C(=O), NO and S(O) p , p is 1 or 2). It includes single-ring, double-ring and tri-ring systems, wherein the double-ring and tri-ring systems include spiro ring, parallel ring and bridged ring. Furthermore, in the case of the "7-10 membered heterocycloalkyl", a heteroatom may occupy the attachment position of the heterocycloalkyl to the rest of the molecule. The 7-10-membered heterocycloalkyl group includes 7-10-membered, 7-9-membered, 7-8-membered, 8-10-membered, 8-9-membered, 8-membered, 9-membered and 10-membered heterocycloalkyl groups, etc. Examples of 7-10 membered heterocycloalkyl groups include, but are not limited to wait.
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选 的实施方式包括但不限于本发明的实施例。The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and methods well known to those skilled in the art. Equivalent replacement method, preferred Implementations include, but are not limited to, embodiments of the present invention.
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：φ/ω扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) uses a Bruker D8 venture diffractometer to collect diffraction intensity data on the cultured single crystal. The light source is CuKα radiation, and the scanning mode is: φ/ω scanning. After collecting relevant data, the direct method is further used. (Shelxs97) The absolute configuration can be confirmed by analyzing the crystal structure.
本发明所使用的溶剂可经市售获得。本发明采用下述缩略词：TMSOTf代表三甲硅基三氟甲磺酸酯；NaBH4代表硼氢化钠；Cs2CO3代表碳酸铯；NBS代表溴代丁二酰亚胺；PE代表石油醚；EtOAc代表乙酸乙酯；DCM代表二氯甲烷；MeOH代表甲醇；EtOH代表乙醇；TFA代表三氟乙酸；Pd(dppf)Cl2代表1,1-双(二苯基磷)二茂铁氯化钯；K2CO3代表碳酸钾；PtO2代表二氧化铂；NaH代表钠氢；THF代表四氢呋喃；Boc代表叔丁基；DMF代表N,N-二甲基甲酰胺；KOH代表氢氧化钾；Xantphos-Pd-G4代表甲烷磺酸[4,5-双二苯基膦-9,9-二甲基氧杂蒽](2-甲基氨基-1,1-联苯-2-基)钯(II)；BrettPhos Pd G3代表甲烷磺酸(2-二环己基膦)-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯)(2'-氨基-1,1'-联苯-2-基)钯(II)；BSA代表牛血清白蛋白；ee代表立体异构体化合物的手性纯度；氟试剂(SelectFlour)试剂代表1-氯甲基-4-氟-1,4-氮鎓双环[2.2.2]辛烷双(四氟硼酸盐)；Pd/C代表钯碳；HOBt代表羟基苯并三唑；DCC代表双环己基酰亚胺；Dess-Martin试剂代表(1,1,1-三乙酰氧基)-1,1-二氢-1,2-苯碘酰-3(1H)-酮。The solvent used in the present invention is commercially available. The present invention adopts the following abbreviations: TMSOTf represents trimethylsilyl triflate; NaBH 4 represents sodium borohydride; Cs 2 CO 3 represents cesium carbonate; NBS represents bromosuccinimide; PE represents petroleum ether ; EtOAc represents ethyl acetate; DCM represents dichloromethane; MeOH represents methanol; EtOH represents ethanol; TFA represents trifluoroacetic acid; Pd(dppf)Cl 2 represents 1,1-bis(diphenylphosphorus)ferrocene chloride Palladium; K 2 CO 3 represents potassium carbonate; PtO 2 represents platinum dioxide; NaH represents sodium hydrogen; THF represents tetrahydrofuran; Boc represents tert-butyl; DMF represents N,N-dimethylformamide; KOH represents potassium hydroxide; Xantphos-Pd-G4 represents palladium methanesulfonate [4,5-bisdiphenylphosphine-9,9-dimethylxanthene](2-methylamino-1,1-biphenyl-2-yl) (II); BrettPhos Pd G3 represents methanesulfonate (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl )(2'-amino-1,1'-biphenyl-2-yl)palladium(II); BSA represents bovine serum albumin; ee represents the chiral purity of stereoisomer compounds; fluorine reagent (SelectFlour) reagent representative 1-Chloromethyl-4-fluoro-1,4-azonium bicyclo[2.2.2]octane bis(tetrafluoroborate); Pd/C represents palladium carbon; HOBt represents hydroxybenzotriazole; DCC represents Dicyclohexyl imide; Dess-Martin reagent represents (1,1,1-triacetoxy)-1,1-dihydro-1,2-phenyliodyl-3(1H)-one.
化合物依据本领域常规命名原则或者使用软件命名，市售化合物采用供应商目录名称。Compounds are named according to conventional naming principles in the field or use For software naming, commercially available compounds adopt supplier catalog names.
技术效果Technical effect
本发明化合物对del19/L858R T790M C797S突变的EGFR展现了较高的激酶抑制活性，且相比野生型具有高选择性，对EGFR三突变细胞具有较好增殖抑制活性，对EGFR野生型增殖抑制活性显著减弱，有较好的选择性。本发明化合物具有良好的体内代谢稳定性，优异的口服吸收药物暴露量和良好的口服吸收生物利用度,在人和小鼠肝微粒体上具有良好的稳定性。The compound of the present invention exhibits high kinase inhibitory activity against del19/L858R T790M C797S mutated EGFR, and is highly selective compared with the wild type. It has good proliferation inhibitory activity against EGFR triple mutant cells and has good proliferation inhibitory activity against EGFR wild type cells. Significantly weakened, with better selectivity. The compound of the invention has good in vivo metabolic stability, excellent oral absorption drug exposure and good oral absorption bioavailability, and has good stability in human and mouse liver microsomes.
附图说明Description of the drawings
图1.化合物A和EGFR突变蛋白的结合模式图；Figure 1. Binding mode diagram of compound A and EGFR mutant protein;
图2.化合物B和EGFR突变蛋白的结合模式图；Figure 2. Binding mode diagram of compound B and EGFR mutant protein;
图3.化合物C和EGFR突变蛋白的结合模式图；Figure 3. Binding mode diagram of compound C and EGFR mutant protein;
图4.化合物D和EGFR突变蛋白的结合模式图；Figure 4. Binding mode diagram of compound D and EGFR mutant protein;
图5.化合物E和EGFR突变蛋白的结合模式图；Figure 5. Binding mode diagram of compound E and EGFR mutant protein;
图6.化合物F和EGFR突变蛋白的结合模式图；Figure 6. Binding mode diagram of compound F and EGFR mutant protein;
图7.化合物G和EGFR突变蛋白的结合模式图；Figure 7. Binding mode diagram of compound G and EGFR mutant protein;
图8.化合物H和EGFR突变蛋白的结合模式图；Figure 8. Binding mode diagram of compound H and EGFR mutant protein;
图9.化合物I和EGFR突变蛋白的结合模式图；Figure 9. Binding mode diagram of compound I and EGFR mutant protein;
图10.化合物J和EGFR突变蛋白的结合模式图。Figure 10. Binding mode diagram of compound J and EGFR mutant protein.
具体实施方式Detailed ways
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。The present invention is described in detail below through examples, which do not mean any adverse limitations to the present invention. The present invention has been described in detail herein, and its specific embodiments are also disclosed. For those skilled in the art, various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. will be obvious.
计算例1
Calculation example 1
分子对接过程是通过使用Maestro(版本2020-1)中的Glide SP[1]和默认选项进行的。选择突变EGFR的共晶体结构(PDB ID：5HCY)作为对接模板。为了准备蛋白质，使用Maestro[2]的蛋白质准备向导模块添加氢原子，并使用OPLS3e力场。对于配体的准备，使用LigPrep模块生成了分子的三维结构，并进行了能量最小化[3]，使用ConfGen模块对小分子构象进行搜索[4]。使用Glide中的Receptor Grid Generation模块生成对接需要的格点(grid)文件，以晶体结构中的配体作为对接盒子的中心。分析蛋白质受体与配体的相互作用类型，然后根据计算得到的对接打分和结合模式选择挑选潜力大的分子进行合成测试。The molecular docking process was performed using Maestro ( Performed with Glide SP[1] in version 2020-1) and default options. The co-crystal structure of mutant EGFR (PDB ID: 5HCY) was selected as the docking template. To prepare the protein, hydrogen atoms were added using the Protein Preparation Wizard module of Maestro [2] and the OPLS3e force field was used. For the preparation of ligands, the LigPrep module was used to generate the three-dimensional structure of the molecule and energy minimization was performed [3], and the ConfGen module was used to search for small molecule conformations [4]. Use the Receptor Grid Generation module in Glide to generate the grid file required for docking, with the ligand in the crystal structure as the center of the docking box. Analyze the interaction type between the protein receptor and the ligand, and then select molecules with high potential for synthesis testing based on the calculated docking score and binding mode selection.
[1]Glide,LLC,New York,NY,2020.[1]Glide, LLC,New York,NY,2020.
[2]Maestro,LLC,New York,NY,2020.[2]Maestro, LLC,New York,NY,2020.
[3]LigPrep,LLC,New York,NY,2020.[3]LigPrep, LLC,New York,NY,2020.
[4]ConfGen,LLC,New York,NY,2020.[4]ConfGen, LLC,New York,NY,2020.
结论：本发明化合物与EGFR突变蛋白有较好的结合。Conclusion: The compound of the present invention binds well to EGFR mutant protein.
中间体1(化合物1h-1、化合物1h-2、化合物1h-3和化合物1h-4)
Intermediate 1 (compound 1h-1, compound 1h-2, compound 1h-3 and compound 1h-4)
步骤1step 1
0℃下，将化合物N-Boc-3-甲基-4-哌啶酮(12g,56.27mmol)和三乙胺(13.66g,135.04mmol)搅拌溶解于甲苯(50mL)中，缓慢滴加TMSOTf(15.01g,67.52mmol)，保持0℃继续搅拌反应4hr。加入60mL水淬灭反应，乙酸乙酯(50mL)萃取两次，合并有机相，并用饱和食盐水洗涤，无水硫酸钠干燥，过滤后减压浓缩得到粗品。经硅胶柱层析(PE:EtOAc＝20:1)纯化得到化合物1d。1H NMR(400MHz,DMSO-d6)δ0.14(s,9H)1.40(s,9H)1.46-1.53(m,3H)2.00-2.09(m,2H)3.38-3.47(m,2H)3.64-3.70(m,2H)。Compound N-Boc-3-methyl-4-piperidone (12g, 56.27mmol) and triethylamine (13.66g, 135.04mmol) were stirred and dissolved in toluene (50mL) at 0°C, and TMSOTf was slowly added dropwise. (15.01g, 67.52mmol), keep stirring at 0°C for 4 hours. The reaction was quenched by adding 60 mL of water, and extracted twice with ethyl acetate (50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. Compound 1d was purified by silica gel column chromatography (PE:EtOAc=20:1). 1 H NMR(400MHz, DMSO-d 6 )δ0.14(s,9H)1.40(s,9H)1.46-1.53(m,3H)2.00-2.09(m,2H)3.38-3.47(m,2H)3.64 -3.70(m,2H).
步骤2Step 2
将化合物1d(10g,35.03mmol)溶解于CH3CN(40mL)，0℃下缓慢加入氟试剂(SelectFlour)试剂(13.65g,38.54mmol)后搅拌反应3hr。反应液加入60mL水淬灭，用乙酸乙酯(3×40mL)萃取后合并有机相，用饱和氯化钠溶液洗涤，无水硫酸钠干燥，过滤后减压浓缩得到化合物1e。1H NMR(400MHz,DMSO-d6)δ1.34-1.38(m,3H)1.42-1.44(m,9H)2.52-2.59(m,2H)3.58-3.86(m,4H)。Compound 1d (10g, 35.03mmol) was dissolved in CH 3 CN (40 mL), fluorine reagent (SelectFlour) reagent (13.65g, 38.54mmol) was slowly added at 0°C, and the reaction was stirred for 3 hours. The reaction solution was quenched by adding 60 mL of water, extracted with ethyl acetate (3 × 40 mL), the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 1e. 1 H NMR (400MHz, DMSO-d 6 ) δ 1.34-1.38 (m, 3H) 1.42-1.44 (m, 9H) 2.52-2.59 (m, 2H) 3.58-3.86 (m, 4H).
步骤3Step 3
将化合物1e(7.2g,31.13mmol)溶解于MeOH(40mL)中，分批缓慢加入NaBH4(1.53g,40.37mmol)后，25℃搅拌反应2hr。加入100mL水淬灭，乙酸乙酯(3×40mL)萃取，合并有机相并用饱和氯化钠溶液洗涤，无水硫酸钠干燥，过滤后浓缩得到化合物1f。1H NMR(400MHz,DMSO-d6)δ1.27(d,J＝1.00Hz,3H)1.39(s,9H)1.49-1.65(m,2H)2.72-2.92(m,2H)3.81-3.95(m,2H)4.98(d,J＝1.00Hz,1H)。Compound 1e (7.2g, 31.13mmol) was dissolved in MeOH (40mL), NaBH 4 (1.53g, 40.37mmol) was slowly added in batches, and the reaction was stirred at 25°C for 2 hr. Add 100 mL of water to quench, extract with ethyl acetate (3 × 40 mL), combine the organic phases and wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter and concentrate to obtain compound 1f. 1 H NMR (400MHz, DMSO-d 6 ) δ1.27 (d, J = 1.00Hz, 3H) 1.39 (s, 9H) 1.49-1.65 (m, 2H) 2.72-2.92 (m, 2H) 3.81-3.95 ( m,2H)4.98(d,J=1.00Hz,1H).
步骤4Step 4
将化合物1f(1g,4.29mmol)溶解于DCM(10mL)中，滴加氯化氢/二氧六环(4M,6.42mL),25℃搅拌反应3hr。将反应液抽滤，收集滤饼即化合物1g的盐酸盐。1H NMR(400MHz,DMSO-d6)δ1.36(d,J＝1.00Hz,3H)1.68-1.86(m,2H)2.84-2.98(m,1H)3.11-3.25(m,2H)3.50-3.61(m,1H)5.18-5.51(m,1H)8.41-8.86(m,1H)9.36-9.59(m,1H)。Compound 1f (1g, 4.29mmol) was dissolved in DCM (10mL), hydrogen chloride/dioxane (4M, 6.42mL) was added dropwise, and the reaction was stirred at 25°C for 3 hr. The reaction solution was suction filtered, and the filter cake, i.e., the hydrochloride salt of 1 g of the compound, was collected. 1 H NMR (400MHz, DMSO-d 6 ) δ1.36 (d, J = 1.00Hz, 3H) 1.68-1.86 (m, 2H) 2.84-2.98 (m, 1H) 3.11-3.25 (m, 2H) 3.50- 3.61(m,1H)5.18-5.51(m,1H)8.41-8.86(m,1H)9.36-9.59(m,1H).
步骤5Step 5
将化合物4-氨基-2-氯嘧啶(3.12g,24.07mmol)和化合物1g的盐酸盐(4.9g,28.89mmol)溶解于异丙醇(50mL)中，加入三乙胺(7.31g,72.22mmol,10.05mL)后加热至100℃搅拌反应16hr。将反应液减压浓缩，经制备HPLC色谱(色谱柱:Phenomenex luna C18 250×80mm×10μm；流动相:[H2O(NH3H2O+NH4HCO3)-乙腈]；乙腈％:0％-20％,20min)分离得产物1h，再经手性制备分离(色谱柱:DAICEL CHIRALCEL OJ(250mm×50mm,10μm)；流动相:超临界CO2-[含0.1％氨水的乙醇溶液]；含0.1％氨水的乙醇溶液％:40％-40％)得到化合物1h-1、1h-2、1h-3和1h-4。分析SFC(色谱柱：ChiralCel OJ-H,150×4.6mm I.D.,5μm；流动相：A(超临界CO2)和B(ethanol(0.05％DEA))；梯度：B％40％)，保留时 间分别为化合物1h-1(Rt＝1.30min)、1h-2(Rt＝1.46min)、1h-3(Rt＝1.72min)和1h-4(Rt＝2.28min)。Compound 4-amino-2-chloropyrimidine (3.12g, 24.07mmol) and the hydrochloride of compound 1g (4.9g, 28.89mmol) were dissolved in isopropanol (50mL), and triethylamine (7.31g, 72.22 mmol, 10.05mL) and then heated to 100°C and stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and subjected to preparative HPLC chromatography (chromatographic column: Phenomenex luna C18 250×80mm×10μm; mobile phase: [H 2 O (NH 3 H 2 O+NH 4 HCO 3 )-acetonitrile]; acetonitrile %: 0%-20%, 20min) to separate the product for 1 hour, and then separate it through chiral preparation (chromatographic column: DAICEL CHIRALCEL OJ (250mm×50mm, 10μm); mobile phase: supercritical CO 2 - [ethanol solution containing 0.1% ammonia] ; Containing 0.1% ammonia in ethanol solution%: 40%-40%) to obtain compounds 1h-1, 1h-2, 1h-3 and 1h-4. Analytical SFC (column: ChiralCel OJ-H, 150×4.6mm ID, 5μm; mobile phase: A (supercritical CO 2 ) and B (ethanol (0.05% DEA)); gradient: B% 40%), retention They are compounds 1h-1 (Rt=1.30min), 1h-2 (Rt=1.46min), 1h-3 (Rt=1.72min) and 1h-4 (Rt=2.28min) respectively.
化合物1h-2:MS m/z:227.2[M+1]+,1H NMR(400MHz,DMSO-d6)δ1.25-1.36(m,3H)1.55-1.68(m,2H)2.81-2.98(m,2H)3.44-3.57(m,1H)4.49-4.59(m,1H)4.62-4.72(m,1H)4.87-5.00(m,1H)5.63-5.77(m,1H)6.25-6.44(m,2H)7.66-7.75(m,1H)。Compound 1h-2:MS m/z:227.2[M+1] + , 1 H NMR (400MHz, DMSO-d 6 )δ1.25-1.36(m,3H)1.55-1.68(m,2H)2.81-2.98 (m,2H)3.44-3.57(m,1H)4.49-4.59(m,1H)4.62-4.72(m,1H)4.87-5.00(m,1H)5.63-5.77(m,1H)6.25-6.44(m ,2H)7.66-7.75(m,1H).
中间体2(化合物17i-1和化合物17i-2)
Intermediate 2 (compound 17i-1 and compound 17i-2)
步骤1step 1
在100mL三口瓶中加入化合物17e(3g,13.81mmol)，四氢呋喃(30mL)，氮气置换三次，降温至-10℃滴入甲基溴化镁(3M,9.21mL)，-10℃搅拌4小时。用饱和氯化铵(10mL)淬灭，然后用乙酸乙酯萃取(20mL×2)，无水硫酸钠干燥，过滤，减压浓缩得到粗品化合物17f。1H NMR(400MHz,CDCl3)δppm 4.15-4.42(m,1H),3.73-4.02(m,1H),3.14-3.68(m,2H),1.72-1.85(m,1H),1.41-1.54(m,12H),1.31(s,3H)。Add compound 17e (3g, 13.81mmol) and tetrahydrofuran (30mL) to a 100mL three-necked flask, replace with nitrogen three times, cool to -10°C, add methylmagnesium bromide (3M, 9.21mL) dropwise, and stir at -10°C for 4 hours. Quenched with saturated ammonium chloride (10 mL), extracted with ethyl acetate (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 17f. 1 H NMR (400MHz, CDCl 3 ) δppm 4.15-4.42(m,1H),3.73-4.02(m,1H),3.14-3.68(m,2H),1.72-1.85(m,1H),1.41-1.54( m,12H),1.31(s,3H).
步骤2Step 2
在反应瓶中加入化合物17f(1.7g,7.29mmol)，氯化氢/二氧六环(4M,17mL)，25℃搅拌2小时。反应液减压浓缩得到粗品化合物17g的盐酸盐。MS m/z:134.2[M+1]+。Compound 17f (1.7g, 7.29mmol) and hydrogen chloride/dioxane (4M, 17mL) were added to the reaction flask, and stirred at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain 17 g of the hydrochloride salt of the crude compound. MS m/z:134.2[M+1] + .
步骤3Step 3
在反应瓶中加入化合物17h(100mg,771.92μmol)，化合物17g(523.74mg,盐酸盐)，二甲基亚砜(5mL)，N,N-二异丙基乙基胺(399.05mg,3.09mmol,537.80μL)，120℃搅拌4小时。反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯(30mL×2)萃取，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品，再通过柱层析纯化(石油醚：乙酸乙酯＝1:0-1:2)得到17i，17i经制备SFC拆分(色谱柱：(DAICEL CHIRALCEL OJ(250mm×30mm,10μm)；流动相：A(超临界CO2)和B(乙醇，含0.1％氨水)；梯度：B％＝25％-25％，10min)得到化合物17i-1和17i-2。Add compound 17h (100 mg, 771.92 μmol), compound 17g (523.74 mg, hydrochloride), dimethyl sulfoxide (5 mL), N, N-diisopropylethylamine (399.05 mg, 3.09) into the reaction bottle. mmol, 537.80 μL), stir at 120°C for 4 hours. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was then passed through the column layer. Analytical purification (petroleum ether: ethyl acetate = 1:0-1:2) gave 17i. 17i was separated by preparative SFC (chromatographic column: (DAICEL CHIRALCEL OJ (250mm×30mm, 10μm)); mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% ammonia); gradient: B% = 25%-25%, 10 min) to obtain compounds 17i-1 and 17i-2.
化合物17j-1：1H NMR(400MHz,DMSO-d6)δppm 7.72(d,J＝5.63Hz,1H),6.40(s,2H),5.71(d,J＝5.63Hz,1H),4.71(s,1H),4.23-4.31(m,1H),4.14(dd,J＝9.01,4.25Hz,1H),3.94-4.06(m,1H),3.38-3.47(m,1H),3.30(s,1H),1.54-1.65(m,1H),1.34-1.49(m,1H),1.19(s,3H)。分析SFC(色谱柱：Chiralcel OJ-3,150×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝10～50％，4min；流速:2.5mL/min；波长:220nm；压力:1800psi)，Rt＝2.832min。Compound 17j-1: 1 H NMR (400MHz, DMSO-d 6 ) δppm 7.72 (d, J = 5.63 Hz, 1H), 6.40 (s, 2H), 5.71 (d, J = 5.63 Hz, 1H), 4.71 ( s,1H),4.23-4.31(m,1H),4.14(dd,J＝9.01,4.25Hz,1H),3.94-4.06(m,1H),3.38-3.47(m,1H),3.30(s, 1H),1.54-1.65(m,1H),1.34-1.49(m,1H),1.19(s,3H). Analytical SFC (chromatographic column: Chiralcel OJ-3, 150×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=10~50%, 4min ; Flow rate: 2.5mL/min; Wavelength: 220nm; Pressure: 1800psi), Rt=2.832min.
化合物17j-2：1H NMR(400MHz,DMSO-d6)δppm 7.72(d,J＝5.63Hz,1H),6.41(s,2H),5.71(d,J＝5.63Hz,1H),4.71(s,1H),4.21-4.34(m,1H),4.14(dd,J＝9.01,4.25Hz,1H),3.89-4.04(m,1H),3.38-3.52(m,1H),3.30(m,1H),1.53-1.67(m,1H),1.30-1.45(m,1H),1.19(s,3H)。分析SFC(色谱柱：Chiralcel OJ-3,150×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝10～50％，4min；流速:2.5mL/min；波长:220nm；压力:1800psi)，Rt＝3.201min。Compound 17j-2: 1 H NMR (400MHz, DMSO-d 6 ) δppm 7.72 (d, J = 5.63 Hz, 1H), 6.41 (s, 2H), 5.71 (d, J = 5.63 Hz, 1H), 4.71 ( s,1H),4.21-4.34(m,1H),4.14(dd,J＝9.01,4.25Hz,1H),3.89-4.04(m,1H),3.38-3.52(m,1H),3.30(m, 1H),1.53-1.67(m,1H),1.30-1.45(m,1H),1.19(s,3H). Analytical SFC (chromatographic column: Chiralcel OJ-3, 150×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=10~50%, 4min ; Flow rate: 2.5mL/min; Wavelength: 220nm; Pressure: 1800psi), Rt=3.201min.
中间体3(化合物23f)
Intermediate 3 (compound 23f)
步骤1 step 1
0℃将NaH(912.10mg,22.80mmol,60％含量)加入至化合物23c(2.5g,11.40mmol)的DMF(25mL)溶液中，搅拌0.5hr，加入碘甲烷(8.09g,57.01mmol,3.55mL)，温度升至25℃搅拌反应16小时。反应液中加入水(300mL)，用乙酸乙酯(3×100mL)萃取，合并有机相并用饱和食盐水洗涤，无水硫酸钠干燥,减压浓缩得到化合物23d。1H NMR(400MHz,CDCl3)δppm 1.48(s,9H)1.75(br s,1H)1.84-1.95(m,1H)3.11(br s,1H)3.23-3.37(m,1H)3.41-3.50(m,4H)3.81(br s,1H)3.99-4.08(m,1H)4.61-4.80(m,1H)。NaH (912.10 mg, 22.80 mmol, 60% content) was added to a solution of compound 23c (2.5 g, 11.40 mmol) in DMF (25 mL) at 0°C, stirred for 0.5 hr, and methyl iodide (8.09 g, 57.01 mmol, 3.55 mL) was added. ), the temperature was raised to 25°C and the reaction was stirred for 16 hours. Water (300 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 × 100 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 23d. 1 H NMR (400MHz, CDCl 3 ) δppm 1.48 (s, 9H) 1.75 (br s, 1H) 1.84-1.95 (m, 1H) 3.11 (br s, 1H) 3.23-3.37 (m, 1H) 3.41-3.50 ( m,4H)3.81(br s,1H)3.99-4.08(m,1H)4.61-4.80(m,1H).
步骤2Step 2
将化合物23d(2.6g,11.15mmol)加入至氯化氢/1,4-二氧六环(4M,25mL)，25℃搅拌2小时。将反应液减压浓缩得到化合物23e。1H NMR(400MHz,DMSO-d6)δppm 1.78-1.87(m,1H)1.89-1.98(m,1H)2.89-3.01(m,1H)3.11-3.30(m,2H)3.32(s,3H)3.35-3.37(m,1H)3.40-3.48(m,1H)3.50-3.64(m,1H)4.98-5.21(m,1H)。Compound 23d (2.6g, 11.15mmol) was added to hydrogen chloride/1,4-dioxane (4M, 25mL), and the mixture was stirred at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain compound 23e. 1 H NMR (400MHz, DMSO-d 6 )δppm 1.78-1.87(m,1H)1.89-1.98(m,1H)2.89-3.01(m,1H)3.11-3.30(m,2H)3.32(s,3H) 3.35-3.37(m,1H)3.40-3.48(m,1H)3.50-3.64(m,1H)4.98-5.21(m,1H).
步骤3Step 3
将化合物23e(1.9g,11.20mmol)，TEA(3.40g,33.60mmol)，化合物17h(1.45g,11.20mmol)加入至异丙醇(30mL)中，100℃搅拌反应16小时。反应完成后直接将反应液减压浓缩，通过制备高效液相色谱(色谱柱:Phenomenex C18 80×40mm×3μm；流动性:[H2O(NH3H2O)-乙腈]；梯度：乙腈％:11％-41％,8min)纯化得到化合物23f。MS m/z:227.1[M+1]+；1H NMR(400MHz,CDCl3)δppm 1.81(dtd,J＝10.01,6.73,6.73,3.89Hz,1H)1.92-2.02(m,1H)3.41-3.48(m,1H)3.48-3.51(m,3H)3.51-3.69(m,2H)4.18-4.27(m,1H)4.46-4.65(m,3H)4.71-4.87(m,1H)5.79(d,J＝5.77Hz,1H)7.95(d,J＝5.52Hz,1H)。Compound 23e (1.9g, 11.20mmol), TEA (3.40g, 33.60mmol), and compound 17h (1.45g, 11.20mmol) were added to isopropanol (30mL), and the reaction was stirred at 100°C for 16 hours. After the reaction is completed, the reaction solution is directly concentrated under reduced pressure and passed through preparative high performance liquid chromatography (chromatographic column: Phenomenex C18 80×40mm×3μm; mobility: [H 2 O (NH 3 H 2 O)-acetonitrile]; gradient: acetonitrile %: 11%-41%, 8 min) and purified to obtain compound 23f. MS m/z: 227.1[M+1] + ; 1 H NMR (400MHz, CDCl 3 ) δppm 1.81 (dtd, J=10.01, 6.73, 6.73, 3.89Hz, 1H) 1.92-2.02 (m, 1H) 3.41- 3.48(m,1H)3.48-3.51(m,3H)3.51-3.69(m,2H)4.18-4.27(m,1H)4.46-4.65(m,3H)4.71-4.87(m,1H)5.79(d, J＝5.77Hz,1H)7.95(d,J＝5.52Hz,1H).
中间体4(化合物28h)
Intermediate 4 (compound 28h)
步骤1step 1
在100mL三口瓶中加入化合物28d(3g,11.84mmol)，二氯甲烷(50mL)，三乙胺(1.44g,14.21mmol,1.98mL)，氮气置换三次，降温至0℃滴入甲烷磺酰氯(1.49g,13.03mmol,1.01mL)，自然升温至25℃搅拌4小时。反应液缓慢加入到冰水溶液(50mL)中淬灭，然后用二氯甲烷萃取(50mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品，经柱层析纯化(石油醚：乙酸乙酯＝1:0-1:1)纯化得到化合物28e。1H NMR(400MHz,CDCl3)δppm 7.32-7.42(m,4H),7.13-7.30(m,6H),4.59(d,J＝6.13Hz,1H),4.38(s,1H),3.71(t,J＝7.32Hz,1H),3.36(t,J＝6.13Hz,1H),2.95(s,3H),2.85(dd,J＝7.94,6.82Hz,1H),0.79(d,J＝6.25Hz,3H)。Add compound 28d (3g, 11.84mmol), dichloromethane (50mL), and triethylamine (1.44g, 14.21mmol, 1.98mL) into a 100mL three-neck flask, replace with nitrogen three times, cool to 0°C and drop in methane sulfonyl chloride ( 1.49g, 13.03mmol, 1.01mL), naturally raise the temperature to 25°C and stir for 4 hours. The reaction solution was slowly added to ice-water solution (50 mL) to quench, and then extracted with dichloromethane (50 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was passed through column layer. Compound 28e was purified by analytical purification (petroleum ether: ethyl acetate = 1:0-1:1). 1 H NMR (400MHz, CDCl 3 ) δppm 7.32-7.42 (m, 4H), 7.13-7.30 (m, 6H), 4.59 (d, J = 6.13Hz, 1H), 4.38 (s, 1H), 3.71 (t ,J＝7.32Hz,1H),3.36(t,J＝6.13Hz,1H),2.95(s,3H),2.85(dd,J＝7.94,6.82Hz,1H),0.79(d,J＝6.25Hz ,3H).
步骤2Step 2
在反应瓶中加入化合物28e(3g,9.05mmol)，甲磺酸乙酸甲酯(1.79g,11.77mmol)，N,N–二甲基甲酰胺(30mL)，氮气置换三次加入氢化钠(432.69mg,10.82mmol,60％)，25℃搅拌15分钟升温至80℃搅拌4小时。反应液缓慢加入到氯化铵水溶液(30mL)中淬灭，然后用乙酸乙酯(50mL×2)萃取，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到产物。粗品通过柱层析纯化(石油醚：乙酸乙酯＝1:0-1:1)纯化得到化合物28f。MS m/z:388.2[M+1]+。Add compound 28e (3g, 9.05mmol), methyl methanesulfonate acetate (1.79g, 11.77mmol), N,N-dimethylformamide (30mL) into the reaction bottle, replace with nitrogen three times and add sodium hydride (432.69mg , 10.82 mmol, 60%), stir at 25°C for 15 minutes, then heat to 80°C and stir for 4 hours. The reaction solution was slowly added to an aqueous ammonium chloride solution (30 mL) to quench, and then extracted with ethyl acetate (50 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 1:0-1:1) to obtain compound 28f. MS m/z:388.2[M+1] + .
步骤3Step 3
在反应瓶中加入化合物28f(1.83g,4.72mmol)，N,N–二甲基乙酰胺(20mL)，氯化锂(1.60g,37.78mmol,773.70μL)氮气置换三次，140℃搅拌10小时。反应液冷却至室温后，反应液缓慢加入到水溶液(30mL)中，然后用乙酸乙酯(50mL×2)萃取，收集有机相，有机相用饱和食盐水(20mL×2)洗涤，无水硫酸钠 干燥，过滤，减压浓缩得到产物。粗品通过柱层析(石油醚：乙酸乙酯＝1:0--1:1)纯化出粗品。粗品经制备高效液相色谱分离(色谱柱:Phenomenex C18 80×40mm×3μm；流动相:A(乙腈)和B(水，含0.05％碳酸氢铵)；梯度：B％:30％-50％，8min)得到化合物28g。1H NMR(400MHz,CDCl3)δppm 7.41(dd,J＝10.07,7.82Hz,4H),7.16-7.32(m,6H),4.33(s,1H),3.66(s,1H),3.04-3.23(m,3H),2.84(s,3H),2.53-2.66(m,2H),0.80(d,J＝6.00Hz,3H)。Add compound 28f (1.83g, 4.72mmol), N,N-dimethylacetamide (20mL), and lithium chloride (1.60g, 37.78mmol, 773.70μL) to the reaction bottle, replace it with nitrogen three times, and stir at 140°C for 10 hours. . After the reaction solution was cooled to room temperature, the reaction solution was slowly added to the aqueous solution (30 mL), and then extracted with ethyl acetate (50 mL × 2). The organic phase was collected, and the organic phase was washed with saturated brine (20 mL × 2) and anhydrous sulfuric acid. sodium Dry, filter and concentrate under reduced pressure to obtain the product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 1:0--1:1). The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Phenomenex C18 80×40mm×3μm; mobile phase: A (acetonitrile) and B (water, containing 0.05% ammonium bicarbonate); gradient: B%: 30%-50% , 8 min) to obtain compound 28g. 1 H NMR (400MHz, CDCl 3 ) δppm 7.41 (dd, J＝10.07, 7.82Hz, 4H), 7.16-7.32 (m, 6H), 4.33 (s, 1H), 3.66 (s, 1H), 3.04-3.23 (m,3H),2.84(s,3H),2.53-2.66(m,2H),0.80(d,J=6.00Hz,3H).
步骤4Step 4
在反应瓶中加入化合物28g(300mg,910.59μmol)，甲醇(5mL)，盐酸(12M,151.77μL)，氢氧化钯(90.00mg,128.17μmol,20％含量)氢气置换三次，25℃,15psi搅拌16小时。反应液进行过滤，滤液减压浓缩得到化合物28h的盐酸盐。1H NMR(400MHz,CD3OD)δppm 4.28(s,1H),3.88(d,J＝9.13Hz,1H),3.72-3.83(m,1H),3.50(dd,J＝9.01,7.75Hz,2H),3.07(dd,J＝7.75Hz,1H),3.00(s,3H),1.52(d,J＝6.50Hz,3H)。Add compound 28g (300mg, 910.59μmol), methanol (5mL), hydrochloric acid (12M, 151.77μL), palladium hydroxide (90.00mg, 128.17μmol, 20% content) into the reaction bottle and replace it with hydrogen three times, stir at 25°C and 15psi 16 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the hydrochloride salt of compound 28h. 1 H NMR (400MHz, CD 3 OD) δppm 4.28 (s, 1H), 3.88 (d, J = 9.13Hz, 1H), 3.72-3.83 (m, 1H), 3.50 (dd, J = 9.01, 7.75Hz, 2H), 3.07 (dd, J = 7.75Hz, 1H), 3.00 (s, 3H), 1.52 (d, J = 6.50Hz, 3H).
实施例1
Example 1
步骤1step 1
向1-Boc-3-(碘甲基)氮杂环丁烷(3.9g,13.13mmol)的乙腈(40mL)/水(13mL)溶液中加入甲硫醇钠(1.86g,26.54mmol)，60℃搅拌18小时。反应液直接浓缩得化合物1a。1H NMR(400MHz,CD3OD)δ3.92-4.06(m,2H),3.58(dd,J＝4.40,8.66Hz,2H),2.61-2.76(m,3H),2.06(s,3H),1.38(s,9H)。To a solution of 1-Boc-3-(iodomethyl)azetidine (3.9g, 13.13mmol) in acetonitrile (40mL)/water (13mL) was added sodium methyl mercaptide (1.86g, 26.54mmol), 60 °C and stirred for 18 hours. The reaction solution was directly concentrated to obtain compound 1a. 1 H NMR (400MHz, CD 3 OD) δ3.92-4.06 (m, 2H), 3.58 (dd, J = 4.40, 8.66Hz, 2H), 2.61-2.76 (m, 3H), 2.06 (s, 3H) ,1.38(s,9H).
步骤2Step 2
向化合物1a(2.1g,9.66mmol)的THF(20mL)/EtOH(20mL)/H2O(20mL)溶液中加入单过硫酸氢钾(8.91g,14.49mmol)，25℃搅拌2小时。向反应液中加水(10mL)，加入乙酸乙酯萃取(5mL×3)，合并有 机相再加入饱和食盐水(10mL)洗涤，有机相用无水硫酸钠干燥，过滤后浓缩得化合物1b。1H NMR(400MHz,CD3OD)δ4.13(t,J＝8.78Hz,2H),3.70-3.83(m,2H),3.29(d,J＝7.54Hz,2H),2.99-3.17(m,1H),2.81-2.92(m,3H),1.39(s,9H)。Potassium monopersulfate (8.91g, 14.49mmol) was added to a solution of compound 1a (2.1g, 9.66mmol) in THF (20mL)/EtOH (20mL)/H 2 O (20mL), and the mixture was stirred at 25°C for 2 hours. Add water (10 mL) to the reaction solution, add ethyl acetate to extract (5 mL × 3), and combine The organic phase was washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 1b. 1 H NMR (400MHz, CD 3 OD) δ4.13 (t, J = 8.78Hz, 2H), 3.70-3.83 (m, 2H), 3.29 (d, J = 7.54Hz, 2H), 2.99-3.17 (m ,1H),2.81-2.92(m,3H),1.39(s,9H).
步骤3Step 3
向化合物1b(2.4g,9.63mmol)的DCM(30mL)溶液中加入三氟乙酸(5.49g,48.13mmol,3.56mL)，25℃搅拌3小时。反应液减压浓缩得化合物1c的三氟乙酸盐。1H NMR(400MHz,DMSO-d6)δ3.97-4.09(m,2H),3.82-3.96(m,2H),3.52(d,J＝7.54Hz,2H),3.24-3.37(m,1H),2.99(s,3H)。To a solution of compound 1b (2.4g, 9.63mmol) in DCM (30mL) was added trifluoroacetic acid (5.49g, 48.13mmol, 3.56mL), and the mixture was stirred at 25°C for 3 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 1c. 1 H NMR (400MHz, DMSO-d 6 ) δ3.97-4.09 (m, 2H), 3.82-3.96 (m, 2H), 3.52 (d, J = 7.54Hz, 2H), 3.24-3.37 (m, 1H ),2.99(s,3H).
步骤4Step 4
向化合物8-溴-3-氯异喹啉(956mg,3.94mmol)，化合物1c(1.04g,3.94mmol,三氟乙酸盐)的1,4-二氧六环(20mL)溶液中加入XantPhos Pd G4(379.39mg,394.23μmol)，Cs2CO3(3.85g,11.83mmol)，氮气保护下100℃搅拌16小时。反应液过滤后直接浓缩得粗品。粗品经薄层色谱法(PE:EtOAc＝1:1)纯化得化合物1i。MS m/z:310.8[M+1]+。To a solution of compound 8-bromo-3-chloroisoquinoline (956 mg, 3.94 mmol), compound 1c (1.04 g, 3.94 mmol, trifluoroacetate salt) in 1,4-dioxane (20 mL) was added XantPhos Pd G4 (379.39mg, 394.23μmol), Cs 2 CO 3 (3.85g, 11.83mmol), stirred at 100°C for 16 hours under nitrogen protection. The reaction solution was filtered and concentrated directly to obtain crude product. The crude product was purified by thin layer chromatography (PE:EtOAc=1:1) to obtain compound 1i. MS m/z:310.8[M+1] + .
步骤5Step 5
向化合物1i(590mg,1.90mmol)的DCM(10mL)溶液中加入NBS(337.87mg,1.90mmol)，25℃搅拌2小时。反应液加入水(10mL)，加入二氯甲烷(5mL×3)萃取，合并有机相并减压浓缩得化合物1j。MS m/z:388.8[M+1]+,390.8[M+3]+。To a solution of compound 1i (590 mg, 1.90 mmol) in DCM (10 mL) was added NBS (337.87 mg, 1.90 mmol), and the mixture was stirred at 25° C. for 2 hours. Water (10 mL) was added to the reaction solution, and dichloromethane (5 mL × 3) was added for extraction. The organic phases were combined and concentrated under reduced pressure to obtain compound 1j. MS m/z:388.8[M+1] + ,390.8[M+3] + .
步骤6Step 6
向化合物1j(850mg,2.18mmol)，异丙烯基硼酸嚬哪醇酯(366.53mg,2.18mmol)的1,4-二氧六环(10mL)/水(2mL)溶液中加入Pd(dppf)Cl2(159.60mg,218.12μmol)，K2CO3(452.19mg,3.27mmol)。氮气保护下65℃搅拌6小时。反应液过滤，减压浓缩得粗品。粗品经硅胶柱层析(PE:EtOAc＝1:1)纯化得化合物1k。MS m/z:350.9[M+1]+。To a solution of compound 1j (850 mg, 2.18 mmol), zonacol isopropenylboronate (366.53 mg, 2.18 mmol) in 1,4-dioxane (10 mL)/water (2 mL) was added Pd(dppf)Cl 2 (159.60 mg, 218.12 μmol), K 2 CO 3 (452.19 mg, 3.27 mmol). Stir at 65°C for 6 hours under nitrogen protection. The reaction solution was filtered and concentrated under reduced pressure to obtain crude product. The crude product was purified by silica gel column chromatography (PE:EtOAc=1:1) to obtain compound 1k. MS m/z:350.9[M+1] + .
步骤7Step 7
向化合物1k(270mg,769.53μmol)的EtOAc(5mL)溶液中加入PtO2(0.5g,2.20mmol)，25℃在氢气球氛围下搅拌2小时。反应液体直接过滤，收集滤液并减压浓缩得化合物1l。MS m/z:352.9[M+1]+。PtO 2 (0.5 g, 2.20 mmol) was added to a solution of compound 1k (270 mg, 769.53 μmol) in EtOAc (5 mL), and the mixture was stirred at 25°C under a hydrogen balloon atmosphere for 2 hours. The reaction liquid was directly filtered, and the filtrate was collected and concentrated under reduced pressure to obtain compound 11. MS m/z:352.9[M+1] + .
步骤8Step 8
向4-吡唑硼酸嚬哪醇酯(2g,10.31mmol)的DMF(20mL)溶液中加入NaH(618.38mg,15.46mmol,60％)，环丙基磺酰氯(1.59g,11.34mmol)，在25℃搅拌5小时。向反应液中加入水(20mL)，加入乙酸乙酯萃取(20mL×3)，合并有机相并用无水硫酸钠干燥，减压浓缩得化合物1m。MS m/z:298.9[M+1]+。To a solution of 4-pyrazoleboronic acid zonacol ester (2g, 10.31mmol) in DMF (20mL), NaH (618.38mg, 15.46mmol, 60%) and cyclopropylsulfonyl chloride (1.59g, 11.34mmol) were added. Stir at 25°C for 5 hours. Water (20 mL) was added to the reaction solution, and ethyl acetate was added for extraction (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 1m. MS m/z:298.9[M+1] + .
步骤9Step 9
向化合物1m(1.54g,5.17mmol)，4-氨基-2-溴嘧啶(0.6g,3.45mmol)的乙腈(15mL)/水(5mL)溶液中加入二氯双[二叔丁基-(4-二甲基氨基苯基)膦]钯(II)(244.17mg,344.83μmol,244.17μL)，Na2CO3(1.10g,10.34mmol)，氮气保护下100℃搅拌3小时。向反应液中加入水(20mL)，加入乙酸乙酯(20mL×3)萃取，合并有机相并用无水硫酸钠干燥，减压浓缩得粗品。粗品经硅胶柱层析(DMC:MeOH＝20:1)纯化得化合物1n。MS m/z:265.9[M+1]+。To a solution of compound 1m (1.54g, 5.17mmol), 4-amino-2-bromopyrimidine (0.6g, 3.45mmol) in acetonitrile (15mL)/water (5mL) was added dichlorobis[di-tert-butyl-(4 -Dimethylaminophenyl)phosphine]palladium(II) (244.17mg, 344.83μmol, 244.17μL), Na 2 CO 3 (1.10g, 10.34mmol), stir at 100°C for 3 hours under nitrogen protection. Water (20 mL) was added to the reaction solution, and ethyl acetate (20 mL × 3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (DMC:MeOH=20:1) to obtain compound 1n. MS m/z:265.9[M+1] + .
步骤10Step 10
向化合物1n(50mg,188.47μmol)，化合物1l(66.51mg,188.47μmol)的1,4-二氧六环(2mL)溶液中加入Xantphos Pd G4(18.14mg,18.85μmol)，Cs2CO3(184.22mg,565.42μmol)，氮气保护下100℃搅拌4小时。向反应液体中加入水(10mL)，加入乙酸乙酯(10mL)萃取，合并有机相，并用无水硫酸钠干燥，减压浓缩得粗品。粗品经制备型HPLC(色谱柱:Welch Xtimate C18 100×40mm×3μm；流动相:[水(三氟乙酸)-乙腈]；乙腈％:30％-60％,8min)分离得化合物1的三氟乙酸盐。MS m/z:582.6[M+1]+；1H NMR(400MHz,DMSO-d6)δ10.55(s,1H),9.12(s,1H),8.75(s,2H),8.67-8.77(m,1H),8.53(s,1H),8.45(d,J＝6.02Hz,1H),7.47 (d,J＝8.04Hz,1H),7.29(br s,1H),6.47(d,J＝8.04Hz,1H),4.42(t,J＝7.78Hz,2H),3.99(t,J＝6.78Hz,2H),3.60(br d,J＝7.54Hz,2H),3.23-3.35(m,2H),3.02(s,3H),1.24-1.40(m,10H)。To a solution of compound 1n (50 mg, 188.47 μmol), compound 1l (66.51 mg, 188.47 μmol) in 1,4-dioxane (2 mL), Xantphos Pd G4 (18.14 mg, 18.85 μmol), Cs 2 CO 3 ( 184.22 mg, 565.42 μmol), stir at 100°C for 4 hours under nitrogen protection. Water (10 mL) was added to the reaction liquid, and ethyl acetate (10 mL) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative HPLC (chromatographic column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile%: 30%-60%, 8min) to obtain the trifluoride of compound 1 acetate. MS m/z:582.6[M+1] + ; 1 H NMR (400MHz, DMSO-d 6 ) δ10.55(s,1H),9.12(s,1H),8.75(s,2H),8.67-8.77 (m,1H),8.53(s,1H),8.45(d,J＝6.02Hz,1H),7.47 (d,J＝8.04Hz,1H),7.29(br s,1H),6.47(d,J＝8.04Hz,1H),4.42(t,J＝7.78Hz,2H),3.99(t,J＝6.78 Hz, 2H), 3.60 (br d, J = 7.54Hz, 2H), 3.23-3.35 (m, 2H), 3.02 (s, 3H), 1.24-1.40 (m, 10H).
实施例2
Example 2
步骤1step 1
0℃将甲基格氏试剂(3.32g,27.88mmol)滴加至N-Boc-氮杂环丁烷-3-甲酸甲酯(2.0g,9.29mmol)的THF(25mL)中，升至室温，反应16hr。滴加饱和氯化铵溶液淬灭反应，用3×15mL乙酸乙酯萃取，合并有机层，减压浓缩得中间体2a。1H NMR(400MHz,DMSO-d6)δ3.72(br s,4H)2.38-2.47(m,1H)1.36(s,9H)0.99(s,6H)。Methyl Grignard reagent (3.32g, 27.88mmol) was added dropwise to N-Boc-azetidine-3-carboxylic acid methyl ester (2.0g, 9.29mmol) in THF (25mL) at 0°C, and the temperature was raised to room temperature. , reaction 16hr. Add saturated ammonium chloride solution dropwise to quench the reaction, extract with 3×15 mL ethyl acetate, combine the organic layers, and concentrate under reduced pressure to obtain intermediate 2a. 1 H NMR (400MHz, DMSO-d 6 ) δ3.72 (br s, 4H) 2.38-2.47 (m, 1H) 1.36 (s, 9H) 0.99 (s, 6H).
步骤2Step 2
0℃将氯化氢/乙酸乙酯(4M,9.29mL)滴加至中间体2a的乙酸乙酯(10mL)中，升至室温，反应16hr。将反应液过滤，滤饼用2mL乙酸乙酯洗涤，干燥得中间体2b的盐酸盐。1H NMR(400MHz,DMSO-d6)δ9.03(br s,1H)8.50(br s,1H)4.93(br s,1H)3.73-3.97(m,4H)2.77(quin,J＝8.47Hz,1H)1.00(s,6H)。Hydrogen chloride/ethyl acetate (4M, 9.29mL) was added dropwise to the ethyl acetate (10mL) of intermediate 2a at 0°C, raised to room temperature, and reacted for 16 hr. The reaction solution was filtered, and the filter cake was washed with 2 mL of ethyl acetate and dried to obtain the hydrochloride salt of intermediate 2b. 1 H NMR (400MHz, DMSO-d 6 ) δ9.03 (br s, 1H) 8.50 (br s, 1H) 4.93 (br s, 1H) 3.73-3.97 (m, 4H) 2.77 (quin, J＝8.47Hz ,1H)1.00(s,6H).
步骤3Step 3
将中间体2b的盐酸盐(0.3g,1.98mmol)，8-溴-3-氯-异喹啉(479.77mg,1.98mmol)，XantPhos PdG4(190.40mg,197.84μmol)，Cs2CO3(1.93g,5.94mmol)加入1,4-二氧六环(15mL)中，氮气保护条件下升温100℃反应8hr。将反应液过滤，浓缩滤液所得粗品经硅胶柱层析(DCM:MeOH＝100:1)得化合物2c。MS m/z:276.9[M+1]+。The hydrochloride salt of intermediate 2b (0.3g, 1.98mmol), 8-bromo-3-chloro-isoquinoline (479.77mg, 1.98mmol), XantPhos PdG4 (190.40mg, 197.84μmol), Cs 2 CO 3 ( 1.93g, 5.94mmol) was added to 1,4-dioxane (15mL), and the temperature was raised to 100°C for 8 hours under nitrogen protection. The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product that was subjected to silica gel column chromatography (DCM:MeOH=100:1) to obtain compound 2c. MS m/z:276.9[M+1] + .
步骤4Step 4
将NBS(257.23mg,1.45mmol)加入含中间体2c(0.4g,1.45mmol)的乙腈(5mL)溶液中，25℃反应2小时。向反应液中加入2mL水淬灭反应，减压蒸除有机溶剂，用2×10mL乙酸乙酯进行萃取，干燥浓缩有机相进行硅胶柱层析(DCM:MeOH＝100:1)得化合物2d。MS m/z:356.8[M+3]+。 NBS (257.23 mg, 1.45 mmol) was added to the acetonitrile (5 mL) solution containing intermediate 2c (0.4 g, 1.45 mmol), and the reaction was carried out at 25°C for 2 hours. Add 2 mL of water to the reaction solution to quench the reaction, evaporate the organic solvent under reduced pressure, extract with 2×10 mL of ethyl acetate, dry and concentrate the organic phase and perform silica gel column chromatography (DCM:MeOH=100:1) to obtain compound 2d. MS m/z:356.8[M+3] + .
步骤5Step 5
氮气保护下，将中间体2d(0.45g,1.27mmol)，2-异丙烯基-4,4,5,5-四甲基-1,3,2-二氧硼酸酯(212.62mg,1.27mmol,237.83μL)，K2CO3(349.74mg,2.53mmol)，Pd(dppf)Cl2(92.58mg,126.53μmol)加入1,4-二氧六环(20mL)/水(4mL)，升温至60℃反应2hr。将反应液倾入20mL水中，用乙酸乙酯(3×8mL)萃取，合并有机相得到中间体2e。MS m/z:316.9[M+1]+。Under nitrogen protection, the intermediate 2d (0.45g, 1.27mmol), 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborate (212.62mg, 1.27 mmol, 237.83μL), K 2 CO 3 (349.74mg, 2.53mmol), Pd(dppf)Cl 2 (92.58mg, 126.53μmol), add 1,4-dioxane (20mL)/water (4mL), and raise the temperature React at 60°C for 2 hours. Pour the reaction solution into 20 mL of water, extract with ethyl acetate (3×8 mL), and combine the organic phases to obtain intermediate 2e. MS m/z:316.9[M+1] + .
步骤6Step 6
向中间体2e(0.2g,631.26μmol)的乙酸乙酯(10mL)溶液，加入PtO2(43.00mg,189.38μmol)，套入氢气球(12.73mg,6.31mmol)，置换后，25℃搅拌反应1hr。将反应液冷却至室温，过滤，浓缩滤液得到中间体2f。MS m/z:319.0[M+1]+。To a solution of intermediate 2e (0.2g, 631.26μmol) in ethyl acetate (10mL), add PtO 2 (43.00mg, 189.38μmol), insert a hydrogen balloon (12.73mg, 6.31mmol), and after replacement, stir the reaction at 25°C. 1hr. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated to obtain intermediate 2f. MS m/z:319.0[M+1] + .
步骤7Step 7
将中间体2f(0.03g,94.09μmol)，中间体1h-2(21.29mg,94.09μmol)，Xantphos Pd G4(9.06mg,9.41μmol)，Cs2CO3(61.31mg,188.18μmol)加入1,4-二氧六环(5mL)，100℃搅拌10小时。将反应液冷却至室温，过滤，经过制备HPLC纯化(色谱柱:Welch Xtimate C18 100×40mm×3μm；流动相:[水(三氟乙酸)-乙腈]；梯度乙腈％:12％-42％,8min)得化合物2。MS m/z:509.3[M+1]+；1H NMR(400MHz,CD3OD-d4)δppm9.25(s,1H)8.25-8.50(m,1H)7.87(d,J＝7.25Hz,1H)7.59(d,J＝8.13Hz,1H)6.70-6.94(m,1H)6.66(d,J＝8.25Hz,1H)4.39-4.51(m,1H)4.24-4.31(m,2H)4.14-4.21(m,2H)3.69-3.82(m,1H)3.50-3.59(m,1H)3.39-3.49(m,1H)3.33(dt,J＝3.22,1.58Hz,2H)2.88-2.99(m,1H)1.97(br dd,J＝9.38,4.00Hz,2H)1.47-1.56(m,3H)1.37(dd,J＝6.82,2.69Hz,6H)1.25(s,6H)。 Add 1 , 4-Dioxane (5 mL), stir at 100°C for 10 hours. The reaction solution was cooled to room temperature, filtered, and purified by preparative HPLC (chromatographic column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient acetonitrile%: 12%-42%, 8min) to obtain compound 2. MS m/z:509.3[M+1] + ; 1 H NMR (400MHz, CD 3 OD-d 4 ) δppm9.25 (s, 1H) 8.25-8.50 (m, 1H) 7.87 (d, J = 7.25Hz ,1H)7.59(d,J＝8.13Hz,1H)6.70-6.94(m,1H)6.66(d,J＝8.25Hz,1H)4.39-4.51(m,1H)4.24-4.31(m,2H)4.14 -4.21(m,2H)3.69-3.82(m,1H)3.50-3.59(m,1H)3.39-3.49(m,1H)3.33(dt,J＝3.22,1.58Hz,2H)2.88-2.99(m, 1H)1.97(br dd,J＝9.38,4.00Hz,2H)1.47-1.56(m,3H)1.37(dd,J＝6.82,2.69Hz,6H)1.25(s,6H).
实施例3
Example 3
步骤1step 1
向6-氯-5-氮杂吲哚(2g,13.11mmol)的DMF(20mL)溶液中加入KOH(2.80g,49.91mmol)，25℃搅拌30分钟，再加入碘单质(3.33g,13.11mmol,2.64mL)加入到上述反应液中，25℃搅拌16小时。向反应液中加入水(50mL)，加入乙酸乙酯(50mL×3)萃取，合并有机相并用饱和食盐水洗涤(100mL)，无水硫酸钠干燥，过滤后滤液减压浓缩得化合物3a。MS m/z:279.1[M+1]+。Add KOH (2.80g, 49.91mmol) to a solution of 6-chloro-5-azaindole (2g, 13.11mmol) in DMF (20mL), stir at 25°C for 30 minutes, and then add iodine element (3.33g, 13.11mmol) , 2.64 mL) was added to the above reaction solution, and stirred at 25°C for 16 hours. Water (50 mL) was added to the reaction solution, and ethyl acetate (50 mL × 3) was added for extraction. The organic phases were combined and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 3a. MS m/z:279.1[M+1] + .
步骤2Step 2
向化合物3a(3.2g,11.49mmol)的DMF(40mL)溶液中，加入NaH(1.38g,34.47mmol,60％纯度)，0℃再加入2-碘丙烷(5.86g,34.47mmol,3.45mL)，25℃搅拌16小时。向反应液中加入水(100mL)，加入乙酸乙酯(60mL×3)萃取，合并有机相并用水洗(100mL)，饱和食盐水洗(100mL)，有机相用无水硫酸钠干燥，过滤，收集滤液减压浓缩得化合物3b。MS m/z:320.8[M+1]+。 To a solution of compound 3a (3.2g, 11.49mmol) in DMF (40mL), add NaH (1.38g, 34.47mmol, 60% purity), and then add 2-iodopropane (5.86g, 34.47mmol, 3.45mL) at 0°C. , stirred at 25°C for 16 hours. Add water (100 mL) to the reaction solution, add ethyl acetate (60 mL × 3) for extraction, combine the organic phases and wash with water (100 mL) and saturated brine (100 mL). The organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is collected. Concentrate under reduced pressure to obtain compound 3b. MS m/z:320.8[M+1] + .
步骤3Step 3
向化合物3b(1.5g,4.68mmol)，化合物1c(1.23g,4.68mmol,三氟乙酸盐)的DMSO(15mL)溶液中加入碘化亚铜(89.12mg,467.94μmol)，L-脯氨酸(107.75mg,935.87μmol)，碳酸钾(2.59g,18.72mmol)。氮气保护下100℃搅拌16小时。向反应液中加水(50mL)，加入乙酸乙酯萃取(50mL×3)萃取，合并有机相并用水洗(60mL×3)，再用食盐水洗涤(60mL×2)，收集有机相并用无水硫酸钠干燥，过滤，滤液减压浓缩得粗品。粗品经硅胶柱层析纯化(PE:EtOAc＝0:1)得化合物3c。MS m/z:341.9[M+1]+。To a solution of compound 3b (1.5g, 4.68mmol) and compound 1c (1.23g, 4.68mmol, trifluoroacetate) in DMSO (15mL), copper iodide (89.12mg, 467.94μmol) and L-proline were added Acid (107.75mg, 935.87μmol), potassium carbonate (2.59g, 18.72mmol). Stir at 100°C for 16 hours under nitrogen protection. Add water (50 mL) to the reaction solution, add ethyl acetate and extract (50 mL Dry over sodium, filter, and concentrate the filtrate under reduced pressure to obtain crude product. The crude product was purified by silica gel column chromatography (PE:EtOAc=0:1) to obtain compound 3c. MS m/z:341.9[M+1] + .
步骤4Step 4
向化合物3c(100mg,292.52μmol)，化合物1h-2(66.18mg,292.52μmol)的1,4-二氧六环(10mL)溶液中加入Xantphos Pd G4(28.15mg,29.25μmol)，Cs2CO3(285.93mg,877.56μmol)，氮气保护下100℃搅拌20小时。反应液过滤，收集滤液减压浓缩得粗品。粗品经薄层色谱法纯化(DCM:MeOH＝10:1)得化合物3。MS m/z:532.2[M+1]+；1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.38(s,1H),8.13(s,1H),7.91(d,J＝5.52Hz,1H),6.68(s,1H),6.34(br d,J＝5.78Hz,1H),5.01(d,J＝6.28Hz,1H),4.62-4.72(m,1H),4.46-4.62(m,2H),4.05-4.16(m,2H),3.64(t,J＝6.78Hz,2H),3.49-3.59(m,2H),3.02-3.27(m,4H),2.98(s,3H),1.69(br s,2H),1.42(d,J＝6.78Hz,6H),1.30-1.39(m,3H)。To a solution of compound 3c (100 mg, 292.52 μmol), compound 1h-2 (66.18 mg, 292.52 μmol) in 1,4-dioxane (10 mL) was added Xantphos Pd G4 (28.15 mg, 29.25 μmol), Cs 2 CO 3 (285.93 mg, 877.56 μmol), stir at 100°C for 20 hours under nitrogen protection. The reaction solution was filtered, and the filtrate was collected and concentrated under reduced pressure to obtain crude product. The crude product was purified by thin layer chromatography (DCM:MeOH=10:1) to obtain compound 3. MS m/z:532.2[M+1] + ; 1 H NMR (400MHz, DMSO-d 6 ) δ9.61(s,1H),8.38(s,1H),8.13(s,1H),7.91(d ,J＝5.52Hz,1H),6.68(s,1H),6.34(br d,J＝5.78Hz,1H),5.01(d,J＝6.28Hz,1H),4.62-4.72(m,1H), 4.46-4.62(m,2H),4.05-4.16(m,2H),3.64(t,J＝6.78Hz,2H),3.49-3.59(m,2H),3.02-3.27(m,4H),2.98( s,3H),1.69(br s,2H),1.42(d,J＝6.78Hz,6H),1.30-1.39(m,3H).
实施例4
Example 4
步骤1step 1
将四氢呋喃(60mL)，二甲基砜(5g,53.12mmol,4.31mL)加入反应瓶中，搅拌；然后将温度降至-65～-70℃后，将正丁基锂(2.5M,21.25mL)滴加到其中后，反应1小时；然后将化合物4a(4.55g,26.56mmol) 溶于四氢呋喃(60mL)中后，滴加到其中，继续反应1小时。向反应体系中加入100mL饱和氯化铵溶液淬灭后，分液；水相用25mL乙酸乙酯萃取后，合并有机相，用饱和食盐水(50mL)洗涤一次后，得到的有机相加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品加入25mL甲基叔丁醚混合搅拌0.5小时后，过滤，得到化合物4b。MS m/z:166.0[M-100+1]+；1H NMR(400MHz,CDCl3)δ＝4.17-4.15(m,2H),4.00-3.96(m,2H),3.64(s,1H),3.48(s,2H),3.06(s,2H),1.45(s,9H)。Add tetrahydrofuran (60mL) and dimethylsulfone (5g, 53.12mmol, 4.31mL) into the reaction flask and stir; then lower the temperature to -65~-70℃, add n-butyllithium (2.5M, 21.25mL ) was added dropwise and allowed to react for 1 hour; then compound 4a (4.55g, 26.56mmol) After being dissolved in tetrahydrofuran (60 mL), it was added dropwise and the reaction was continued for 1 hour. Add 100 mL of saturated ammonium chloride solution to the reaction system to quench and separate the liquids; extract the aqueous phase with 25 mL of ethyl acetate, combine the organic phases, wash once with saturated brine (50 mL), and add anhydrous to the obtained organic phase. After drying over sodium sulfate, it was filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. 25 mL of methyl tert-butyl ether was added to the crude product, mixed and stirred for 0.5 hours, and then filtered to obtain compound 4b. MS m/z: 166.0 [M-100+1] + ; 1 H NMR (400MHz, CDCl 3 ) δ = 4.17-4.15 (m, 2H), 4.00-3.96 (m, 2H), 3.64 (s, 1H) ,3.48(s,2H),3.06(s,2H),1.45(s,9H).
步骤2Step 2
将乙酸乙酯(5mL)，化合物4b(0.5g,1.88mmol)加入反应瓶中，开始搅拌；然后将温度降至0～5℃后，将氯化氢/乙酸乙酯(4M,10mL)加入其中，升至室温18℃，反应2小时。反应液直接过滤，滤饼真空干燥得到化合物4c的盐酸盐。1H NMR(400MHz,CD3OD)δ4.39-4.36(m,2H),4.06-4.01(m,2H),3.86-3.66(m,3H),3.07(s,2H)。Add ethyl acetate (5mL) and compound 4b (0.5g, 1.88mmol) into the reaction flask and start stirring; then, after the temperature is lowered to 0~5°C, hydrogen chloride/ethyl acetate (4M, 10mL) is added, Raise to room temperature 18°C and react for 2 hours. The reaction solution was directly filtered, and the filter cake was dried under vacuum to obtain the hydrochloride of compound 4c. 1 H NMR (400MHz, CD 3 OD) δ4.39-4.36(m,2H), 4.06-4.01(m,2H), 3.86-3.66(m,3H), 3.07(s,2H).
步骤3Step 3
在反应瓶中加入化合物4d(2.3g,12.74mmol)，N,N-二甲基甲酰胺(20mL)，氮气置换三次降温至0℃加入N-碘代丁二酰亚胺(3.44g,15.28mmol)升温至25℃搅拌24小时。反应完成后，反应液进行过滤，滤饼用水(10mL)洗涤，收集滤饼，滤饼真空干燥得到化合物4e。1H NMR(400MHz,DMSO-d6)δppm 11.96-12.10(m,1H),9.03-9.11(m,1H),7.91(s,1H),7.48(s,1H)。Add compound 4d (2.3g, 12.74mmol) and N,N-dimethylformamide (20mL) to the reaction flask, replace with nitrogen three times and cool to 0°C. Add N-iodosuccinimide (3.44g, 15.28 mmol) was heated to 25°C and stirred for 24 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with water (10 mL), the filter cake was collected, and the filter cake was dried under vacuum to obtain compound 4e. 1 H NMR (400MHz, DMSO-d 6 ) δ ppm 11.96-12.10 (m, 1H), 9.03-9.11 (m, 1H), 7.91 (s, 1H), 7.48 (s, 1H).
步骤4Step 4
在反应瓶中加入化合物4e(2.2g,7.18mmol)，氧氯化磷(16mL)，氮气置换三次，100℃搅拌5小时。反应完成后将反应液进行减压浓缩得到粗品，粗品使用乙酸乙酯(50mL)溶解，将溶剂中的粗品加入到碳酸氢钠水溶液中淬灭，使用乙酸乙酯萃取(20mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，滤液减压浓缩，用硅胶柱层析纯化(石油醚：乙酸乙酯＝100:1～5:1)纯化得到化合物4f。1H NMR(400MHz,DMSO-d6)δppm 9.34-9.50(m,1H),8.91-9.07(m,1H),7.77-8.00(m,1H)。Compound 4e (2.2g, 7.18mmol) and phosphorus oxychloride (16mL) were added to the reaction flask, replaced with nitrogen three times, and stirred at 100°C for 5 hours. After the reaction is completed, the reaction solution is concentrated under reduced pressure to obtain a crude product. The crude product is dissolved in ethyl acetate (50 mL). The crude product in the solvent is added to an aqueous sodium bicarbonate solution to quench, and extracted with ethyl acetate (20 mL × 2). Collect. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1 to 5:1) to obtain compound 4f. 1 H NMR (400MHz, DMSO-d 6 ) δ ppm 9.34-9.50 (m, 1H), 8.91-9.07 (m, 1H), 7.77-8.00 (m, 1H).
步骤5Step 5
在反应瓶中加入化合物4f(1g,3.08mmol)，异丙烯基硼酸嚬哪醇酯(620.59mg,3.69mmol)，碳酸钾(1.28g,9.23mmol)，二氧六环(10mL)，水(2mL)，氮气置换三次加入二氯双[二叔丁基-(4-二甲基氨基苯基)膦]钯(II)(435.83mg,615.51μmol)，50℃搅拌2小时，反应完成后，反应液冷却至室温后加入水(10mL)后用乙酸乙酯萃取(20mL×3)，收集有机相，有机相用饱和食盐水洗涤(10mL×2)，无水硫酸钠干燥，过滤，滤液减压浓缩得到粗品。粗品用硅胶柱层析纯化(石油醚：乙酸乙酯＝100:0～5:1)纯化得到化合物4g。1H NMR(400MHz,DMSO-d6)δppm 9.49-9.78(m,1H),8.38-8.55(m,1H),7.91-8.09(m,1H),5.55-5.69(m,1H),5.10-5.27(m,1H),2.17-2.22(m,3H)。Add compound 4f (1g, 3.08mmol), zonacol isopropenyl borate (620.59mg, 3.69mmol), potassium carbonate (1.28g, 9.23mmol), dioxane (10mL), water ( 2 mL), replaced with nitrogen three times, added dichlorobis[di-tert-butyl-(4-dimethylaminophenyl)phosphine]palladium(II) (435.83 mg, 615.51 μmol), and stirred at 50°C for 2 hours. After the reaction was completed, After the reaction solution was cooled to room temperature, water (10 mL) was added and extracted with ethyl acetate (20 mL × 3). The organic phase was collected, washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was reduced to Concentrate under pressure to obtain crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:0~5:1) to obtain 4 g of compound. 1 H NMR (400MHz, DMSO-d 6 ) δppm 9.49-9.78(m,1H),8.38-8.55(m,1H),7.91-8.09(m,1H),5.55-5.69(m,1H),5.10- 5.27(m,1H),2.17-2.22(m,3H).
步骤6Step 6
在反应瓶中加入化合物4g(900mg,3.76mmol)，乙酸乙酯(20mL)，二氧化铂(1.11g,4.89mmol)，氢气球置换三次氢气，25℃，15psi搅拌16小时。反应液过滤，滤饼用乙酸乙酯洗涤(20mL)，合并滤液，减压浓缩得到粗产品。再用硅胶柱层析纯化(石油醚：乙酸乙酯＝100:0～5:1)得到化合物4h。MS m/z:241.0[M+1]+。1H NMR(400MHz,DMSO-d6)δppm 9.46-9.56(m,1H)8.46-8.55(m,1H)8.19-8.36(m,1H)3.54-3.76(m,1H)1.31-1.37(m,6H)。Add compound 4g (900mg, 3.76mmol), ethyl acetate (20mL), and platinum dioxide (1.11g, 4.89mmol) into the reaction bottle. Replace the hydrogen gas with a hydrogen balloon three times. Stir at 25°C and 15psi for 16 hours. The reaction solution was filtered, and the filter cake was washed with ethyl acetate (20 mL). The filtrate was combined and concentrated under reduced pressure to obtain a crude product. Then, it was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:0~5:1) to obtain compound 4h. MS m/z:241.0[M+1] + . 1 H NMR(400MHz, DMSO-d 6 )δppm 9.46-9.56(m,1H)8.46-8.55(m,1H)8.19-8.36(m,1H)3.54-3.76(m,1H)1.31-1.37(m, 6H).
步骤7Step 7
将异丙醇(4mL)，化合物4c(125.46mg,622.11μmol,盐酸盐)加入反应瓶中，开始搅拌；然后将化合物4h(100mg,414.74μmol)，三乙胺(146.89mg,1.45mmol,202.04μL)加入其中，升温至100℃，反应2小时。向反应液中加入15mL饱和氯化铵溶液以及15mL乙酸乙酯分液；得到的水相用15mL乙酸乙酯萃取后，合并有机相，用饱和食盐水15mL洗涤后，加入无水硫酸钠干燥，过滤，滤液在50℃下进行减压浓缩。粗品经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝5:1～1:4)。得到化合物4i。MS m/z:370.0[M+1]+； 1H NMR(400MHz,DMSO-d6)δ＝9.07(s,1H),8.14(s,1H),7.89(s,1H),6.54(s,1H),4.61(d,J＝9.6Hz,2H),4.31(d,J＝9.6Hz,2H),3.69(s,2H),3.42-3.35(m,1H),3.04(s,3H),1.27(d,J＝6.8Hz,6H)。Add isopropyl alcohol (4mL) and compound 4c (125.46mg, 622.11μmol, hydrochloride) into the reaction bottle and start stirring; then add compound 4h (100mg, 414.74μmol), triethylamine (146.89mg, 1.45mmol, 202.04 μL) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. Add 15 mL of saturated ammonium chloride solution and 15 mL of ethyl acetate to the reaction solution for separation; extract the obtained aqueous phase with 15 mL of ethyl acetate, combine the organic phases, wash with 15 mL of saturated brine, and dry by adding anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure at 50°C. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1~1:4). Compound 4i was obtained. MS m/z:370.0[M+1] + ; 1 H NMR (400MHz, DMSO-d 6 ) δ = 9.07 (s, 1H), 8.14 (s, 1H), 7.89 (s, 1H), 6.54 (s, 1H), 4.61 (d, J = 9.6Hz, 2H), 4.31 (d, J = 9.6Hz, 2H), 3.69 (s, 2H), 3.42-3.35 (m, 1H), 3.04 (s, 3H), 1.27 (d, J = 6.8Hz, 6H).
步骤8Step 8
将1,4-二氧六环(5mL)，化合物4i(50mg,135.18μmol)加入反应瓶中，开始搅拌；然后将碳酸铯(88.09mg,270.37μmol)，化合物1h-2(33.64mg,148.70μmol)加入其中，氮气置换后将BrettPhos Pd G3(36.76mg,40.56μmol)加入其中，升温至100℃，反应5小时。向反应体系中加入15mL饱和氯化铵溶液以及15mL乙酸乙酯；得到的水相用15mL乙酸乙酯萃取后，合并有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：二氯甲烷:甲醇＝100:0～90:10)，再用1.5mL的甲基叔丁基醚/二氯甲烷(4:1)混合液搅拌15min后，过滤，干燥得到化合物4。MS m/z:560.3[M+1]+；1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),9.00(s,1H),8.49(s,1H),8.04-7.99(m,2H),6.50(s,2H),5.04(d,J＝6.4Hz,1H),4.76-4.64(m,2H),4.59(d,J＝9.2Hz,2H),4.28(d,J＝8.8Hz,2H),3.70(s,2H),3.61-3.50(m,1H),3.22-3.05(m,5H),1.73(s,2H),1.39-1.30(m,9H)。Add 1,4-dioxane (5mL), compound 4i (50mg, 135.18μmol) into the reaction bottle and start stirring; then add cesium carbonate (88.09mg, 270.37μmol), compound 1h-2 (33.64mg, 148.70 μmol) was added, and after nitrogen replacement, BrettPhos Pd G3 (36.76 mg, 40.56 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 5 hours. Add 15 mL of saturated ammonium chloride solution and 15 mL of ethyl acetate to the reaction system; extract the obtained aqueous phase with 15 mL of ethyl acetate, wash the combined organic phases with 15 mL of saturated brine, add anhydrous sodium sulfate, dry, and filter. The filtrate was concentrated under reduced pressure at 45°C to obtain crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution: dichloromethane: methanol = 100:0~90:10), and then 1.5 mL of methyl tert-butyl ether/dichloromethane (4:1) mixture was used After stirring for 15 min, it was filtered and dried to obtain compound 4. MS m/z:560.3[M+1] + ; 1 H NMR (400MHz, DMSO-d 6 ) δ10.09(s,1H),9.00(s,1H),8.49(s,1H),8.04-7.99 (m,2H),6.50(s,2H),5.04(d,J＝6.4Hz,1H),4.76-4.64(m,2H),4.59(d,J＝9.2Hz,2H),4.28(d, J＝8.8Hz,2H),3.70(s,2H),3.61-3.50(m,1H),3.22-3.05(m,5H),1.73(s,2H),1.39-1.30(m,9H).
实施例5
Example 5
步骤1step 1
将三乙胺(19.05g,188.24mmol,26.20mL)加入到含化合物5a(30g,125.36mmol)的DCM(300mL)溶液中，冷却至-20℃，再将甲烷磺酰氯(17.76g,155.04mmol,12.00mL)滴加至反应液中，升至0℃搅拌1小时。将反应液缓慢到入冰水中(300mL)，分层收集有机相，再用10％的碳酸氢钠溶液洗涤，有机相用 无水硫酸钠干燥，过滤后滤液减压浓缩得化合物5b。MS m/z:318.1[M+1]+。Triethylamine (19.05g, 188.24mmol, 26.20mL) was added to a solution of compound 5a (30g, 125.36mmol) in DCM (300mL), cooled to -20°C, and then methanesulfonyl chloride (17.76g, 155.04mmol) , 12.00 mL) was added dropwise to the reaction solution, raised to 0°C and stirred for 1 hour. The reaction solution was slowly poured into ice water (300 mL), the organic phase was collected in layers, and then washed with 10% sodium bicarbonate solution. Dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain compound 5b. MS m/z:318.1[M+1] + .
步骤2Step 2
0℃将丙二酸二乙酯(28.86g,180.21mmol,27.23mL)滴加至钠氢(6.54g,163.50mmol,60％)的N,N-二甲基甲酰胺(180mL)溶液中，低于25℃搅拌1小时，再加入化合物5b(26g,81.92mmol)的N,N-二甲基甲酰胺(40mL)，70℃搅拌16小时。将反应液体加入到冰水中(400mL)，加入乙酸乙酯萃取(200mL×3)，合并有机相，再用水洗(300mL×3)，在用饱和氯化钠水溶液萃取(300mL)，有机相用无水硫酸钠干燥，减压浓缩得粗品。粗品经硅胶柱层析分离(PE:EA＝5:1)得化合物5c。MS m/z:382.0[M+1]+；1H NMR(400MHz,CDCl3)δ7.38(d,J＝7.28Hz,4H),7.24(t,J＝7.54Hz,4H),7.10-7.19(m,2H),4.31(s,1H),4.07-4.17(m,4H),3.64(d,J＝10.54Hz,1H),3.30-3.41(m,2H),2.96-3.09(m,1H),2.85-2.94(m,2H),1.17-1.24(m,6H)。Diethyl malonate (28.86g, 180.21mmol, 27.23mL) was added dropwise to a solution of sodium hydrogen (6.54g, 163.50mmol, 60%) in N,N-dimethylformamide (180mL) at 0°C. Stir below 25°C for 1 hour, then add compound 5b (26g, 81.92mmol) in N,N-dimethylformamide (40mL), and stir at 70°C for 16 hours. Add the reaction liquid to ice water (400mL), add ethyl acetate for extraction (200mL×3), combine the organic phases, wash with water (300mL×3), and extract with saturated sodium chloride aqueous solution (300mL). The organic phase is Dry over anhydrous sodium sulfate and concentrate under reduced pressure to obtain crude product. The crude product was separated by silica gel column chromatography (PE:EA=5:1) to obtain compound 5c. MS m/z: 382.0[M+1] + ; 1 H NMR (400MHz, CDCl 3 ) δ7.38 (d, J = 7.28Hz, 4H), 7.24 (t, J = 7.54Hz, 4H), 7.10- 7.19(m,2H),4.31(s,1H),4.07-4.17(m,4H),3.64(d,J＝10.54Hz,1H),3.30-3.41(m,2H),2.96-3.09(m, 1H),2.85-2.94(m,2H),1.17-1.24(m,6H).
步骤3Step 3
15℃向含四氢锂铝(2.39g,63.01mmol)的四氢呋喃(125mL)混悬液中加入化合物5c(10g,26.21mmol)，15℃搅拌16小时。向反应液中加入缓慢水(2.4mL)，再加入15％的氢氧化钠水溶液(2.4mL)，水(7.2mL)，继续搅拌30分钟，过滤后滤液减压浓缩得化合物5d。1H NMR(400MHz,CDCl3)δ7.39(br d,J＝7.28Hz,4H),7.29(t,J＝7.40Hz,4H),7.15-7.24(m,2H),4.35(s,1H),4.03-4.27(m,1H),3.54-3.80(m,4H),3.23-3.49(m,3H),2.86-3.01(m,2H),2.38-2.58(m,1H),1.85-2.02(m,1H)。Compound 5c (10 g, 26.21 mmol) was added to a suspension of tetrahydrofuran (125 mL) containing lithium aluminum tetrahydrogen (2.39 g, 63.01 mmol) at 15° C., and the mixture was stirred at 15° C. for 16 hours. Slowly add water (2.4 mL) to the reaction solution, then add 15% sodium hydroxide aqueous solution (2.4 mL) and water (7.2 mL), continue stirring for 30 minutes, filter and concentrate the filtrate under reduced pressure to obtain compound 5d. 1 H NMR (400MHz, CDCl 3 ) δ7.39 (br d, J＝7.28Hz, 4H), 7.29 (t, J＝7.40Hz, 4H), 7.15-7.24 (m, 2H), 4.35 (s, 1H ),4.03-4.27(m,1H),3.54-3.80(m,4H),3.23-3.49(m,3H),2.86-3.01(m,2H),2.38-2.58(m,1H),1.85-2.02 (m,1H).
步骤4Step 4
将湿Pd/C(3.5g,23.54mmol,10％)加入到含化合物5d(7g,23.54mmol)，Boc酸酐(7.71g,35.31mmol)的乙酸乙酯(150mL)溶液中，反应液在45psi条件下氢化，25℃搅拌36小时。反应液过滤，收集滤液，减压浓缩得粗品。粗品经硅胶柱层析(DCM:MeOH＝10:1)分离得化合物5e。1H NMR(400MHz,CDCl3)δ3.99(t,J＝8.54Hz,2H),3.67-3.85(m,4H),3.61(dd,J＝6.54,10.80Hz,2H),2.53-2.66(m,1H),1.85-1.96(m,1H),1.42(s,9H)。Wet Pd/C (3.5g, 23.54mmol, 10%) was added to a solution of compound 5d (7g, 23.54mmol) and Boc anhydride (7.71g, 35.31mmol) in ethyl acetate (150mL). The reaction solution was maintained at 45psi. Hydrogenate under normal conditions and stir at 25°C for 36 hours. The reaction solution was filtered, the filtrate was collected, and concentrated under reduced pressure to obtain crude product. The crude product was separated by silica gel column chromatography (DCM:MeOH=10:1) to obtain compound 5e. 1 H NMR (400MHz, CDCl 3 ) δ3.99 (t, J = 8.54Hz, 2H), 3.67-3.85 (m, 4H), 3.61 (dd, J = 6.54, 10.80Hz, 2H), 2.53-2.66 ( m,1H),1.85-1.96(m,1H),1.42(s,9H).
步骤5Step 5
0℃向含有三苯基膦(3.40g,12.97mmol)的二氯甲烷(20mL)中滴加液溴(2.07g,12.97mmol,0.67mL)，0℃搅拌1小时，再滴加三乙胺(1.31g,12.97mmol,1.81mL)，0℃继续搅拌1小时，再滴加化合物5e(1g,4.32mmol)的二氯甲烷溶液，25℃搅拌16小时。反应液加水(100mL)，加入二氯甲烷(100mL)萃取，有机相减压浓缩得粗品。粗品经硅胶柱层析(PE:EA＝5:1)分离得化合物5f。1H NMR(400MHz,CDCl3)δ4.03(t,J＝8.54Hz,2H),3.70(dd,J＝6.02,8.78Hz,2H),3.57(dd,J＝3.40,10.42Hz,2H),3.42(dd,J＝6.54,10.54Hz,2H),2.66(ttd,J＝6.06,8.26,10.52Hz,1H),2.23(ttd,J＝3.42,6.69,10.15Hz,1H),1.43(s,9H)。Add liquid bromine (2.07g, 12.97mmol, 0.67mL) dropwise to methylene chloride (20mL) containing triphenylphosphine (3.40g, 12.97mmol) at 0°C, stir at 0°C for 1 hour, and then add triethylamine dropwise. (1.31g, 12.97mmol, 1.81mL), continue stirring at 0°C for 1 hour, then add dropwise a dichloromethane solution of compound 5e (1g, 4.32mmol), and stir at 25°C for 16 hours. Water (100 mL) was added to the reaction solution, and dichloromethane (100 mL) was added for extraction. The organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (PE:EA=5:1) to obtain compound 5f. 1 H NMR (400MHz, CDCl 3 ) δ4.03 (t, J = 8.54Hz, 2H), 3.70 (dd, J = 6.02, 8.78Hz, 2H), 3.57 (dd, J = 3.40, 10.42Hz, 2H) ,3.42(dd,J＝6.54,10.54Hz,2H),2.66(ttd,J＝6.06,8.26,10.52Hz,1H),2.23(ttd,J＝3.42,6.69,10.15Hz,1H),1.43(s ,9H).
步骤6Step 6
向化合物5f(1.08g,3.02mmol)的乙腈(25mL)/水(2.5mL)混合溶液中加入硫化钠(944.19mg,12.10mmol)，50℃搅拌12小时。向反应液中加入水(50mL)，加入二氯甲烷(50mL×3)萃取，合并有机相再用饱和碳酸氢钠洗涤，有机相用无水硫酸钠干燥，减压浓缩得化合物5g。1H NMR(400MHz,CDCl3)δ3.98(br t,J＝8.38Hz,2H),3.51-3.72(m,2H),3.45(qd,J＝7.78,15.70Hz,1H),3.22(br t,J＝8.88Hz,2H),2.88-3.00(m,2H),2.67-2.85(m,1H),1.42(s,9H)。Sodium sulfide (944.19 mg, 12.10 mmol) was added to a mixed solution of compound 5f (1.08 g, 3.02 mmol) in acetonitrile (25 mL)/water (2.5 mL), and the mixture was stirred at 50° C. for 12 hours. Water (50 mL) was added to the reaction solution, and dichloromethane (50 mL × 3) was added for extraction. The organic phases were combined and washed with saturated sodium bicarbonate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 5 g of the compound. 1 H NMR (400MHz, CDCl 3 ) δ3.98 (br t, J＝8.38Hz, 2H), 3.51-3.72 (m, 2H), 3.45 (qd, J＝7.78, 15.70Hz, 1H), 3.22 (br t,J＝8.88Hz,2H),2.88-3.00(m,2H),2.67-2.85(m,1H),1.42(s,9H).
步骤7Step 7
0℃向化合物5g(0.68g,2.97mmol)的二氯甲烷(15mL)溶液中加入间氯过氧苯甲酸(1.26g,6.23mmol,85％含量)，缓慢升至25℃搅拌16小时。反应液直接过滤，收集滤液，滤液加入饱和硫代硫酸钠溶液(100mL)洗涤，再接着用10％的碳酸钠水溶液(100mL)洗涤，最后用水(100mL)洗涤，有机相用无水硫酸钠干燥，过滤，滤液减压浓缩得化合物5h。MS m/z:205.8[M-56+1]+；1H NMR(400MHz,CDCl3)δ4.17-4.35(m,2H),4.08(t,J＝8.38Hz,2H),3.70-3.87(m,2H),3.57(br dd,J＝3.94,8.70Hz,2H),2.73-2.95(m,2H),1.43(s,9H)。To a solution of 5 g of compound (0.68 g, 2.97 mmol) in dichloromethane (15 mL) was added m-chloroperoxybenzoic acid (1.26 g, 6.23 mmol, 85% content) at 0° C., and the solution was slowly raised to 25° C. and stirred for 16 hours. The reaction solution was directly filtered, and the filtrate was collected. The filtrate was washed with saturated sodium thiosulfate solution (100mL), then washed with 10% sodium carbonate aqueous solution (100mL), and finally washed with water (100mL). The organic phase was dried with anhydrous sodium sulfate. , filtered, and the filtrate was concentrated under reduced pressure to obtain compound 5h. MS m/z: 205.8[M-56+1] + ; 1 H NMR (400MHz, CDCl 3 ) δ4.17-4.35 (m, 2H), 4.08 (t, J = 8.38Hz, 2H), 3.70-3.87 (m,2H),3.57(br dd,J＝3.94,8.70Hz,2H),2.73-2.95(m,2H),1.43(s,9H).
步骤8 Step 8
0℃向化合物5h(0.1g,382.65μmol)的DCM(2mL)溶液中加入三氟乙酸(775.75mg,6.80mmol,503.73μL)，25℃搅拌16小时。反应液减压浓缩得化合物5i的三氟乙酸盐。1H NMR(400MHz,CDCl3)δ7.81-9.30(m,1H),4.31(br s,4H),3.62-4.08(m,4H),2.92-3.55(m,2H)。Trifluoroacetic acid (775.75 mg, 6.80 mmol, 503.73 μL) was added to a solution of compound 5h (0.1 g, 382.65 μmol) in DCM (2 mL) at 0°C, and the mixture was stirred at 25°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 5i. 1 H NMR (400MHz, CDCl 3 ) δ7.81-9.30 (m, 1H), 4.31 (br s, 4H), 3.62-4.08 (m, 4H), 2.92-3.55 (m, 2H).
步骤9Step 9
向化合物4h(40mg,165.90μmol)，化合物5i(59.36mg,215.67μmol,三氟乙酸盐)的异丙醇(2mL)溶液中加入三乙胺(67.15mg,663.60μmol,92.36μL)，100℃搅拌2小时。反应液浓缩直接薄层色谱法纯化(DCM:MeOH＝20:1)得化合物5j。MS m/z:365.9[M+1]+。To a solution of compound 4h (40 mg, 165.90 μmol) and compound 5i (59.36 mg, 215.67 μmol, trifluoroacetate) in isopropanol (2 mL), triethylamine (67.15 mg, 663.60 μmol, 92.36 μL) was added, 100 °C and stir for 2 hours. The reaction solution was concentrated and directly purified by thin layer chromatography (DCM:MeOH=20:1) to obtain compound 5j. MS m/z:365.9[M+1] + .
步骤10Step 10
向化合物5j(20mg,54.66μmol)，化合物1h-2(12.37mg,54.66μmol)的1,4-二氧六环(2mL)溶液中加入BrettPhos Pd-G4(5.03mg,5.47μmol)，BrettPhos Pd-G3(4.96mg,5.47μmol)，Cs2CO3(71.24mg,218.65μmol)。氮气保护下100℃搅拌16小时。反应液过滤，滤液浓缩得粗品。粗品经制备分离板(DCM:MeOH＝10:1)分离得粗品，继续经过Pre-HPLC纯化(色谱柱:Welch Xtimate C18 100×40mm×3μm；流动相:[水(三氟乙酸)-乙腈]；梯度：乙腈％:0％-28％,8min)，得化合物5的三氟乙酸盐。MS m/z:556.1[M+1]+；1H NMR(400MHz,CD3OD)δ9.28(s,1H),8.40(br s,1H),8.04(d,J＝7.04Hz,1H),7.46(s,1H),7.04(br s,1H),4.93(br d,J＝6.54Hz,2H),4.55(br s,4H),4.38(br dd,J＝10.04,14.06Hz,2H),4.03(br dd,J＝5.90,14.44Hz,2H),3.65-3.84(m,1H),3.34-3.53(m,4H),3.03-3.18(m,1H),1.95(br s,2H),1.45-1.60(m,3H),1.39(d,J＝6.78Hz,6H)。To a solution of compound 5j (20 mg, 54.66 μmol), compound 1h-2 (12.37 mg, 54.66 μmol) in 1,4-dioxane (2 mL) was added BrettPhos Pd-G4 (5.03 mg, 5.47 μmol), BrettPhos Pd -G3 (4.96 mg, 5.47 μmol), Cs 2 CO 3 (71.24 mg, 218.65 μmol). Stir at 100°C for 16 hours under nitrogen protection. The reaction solution was filtered, and the filtrate was concentrated to obtain crude product. The crude product was separated by a preparative separation plate (DCM:MeOH=10:1) and continued to be purified by Pre-HPLC (chromatographic column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile] ; Gradient: acetonitrile%: 0%-28%, 8min), the trifluoroacetate salt of compound 5 was obtained. MS m/z:556.1[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δ9.28 (s, 1H), 8.40 (br s, 1H), 8.04 (d, J = 7.04Hz, 1H ),7.46(s,1H),7.04(br s,1H),4.93(br d,J＝6.54Hz,2H),4.55(br s,4H),4.38(br dd,J＝10.04,14.06Hz, 2H),4.03(br dd,J＝5.90,14.44Hz,2H),3.65-3.84(m,1H),3.34-3.53(m,4H),3.03-3.18(m,1H),1.95(br s, 2H), 1.45-1.60 (m, 3H), 1.39 (d, J = 6.78Hz, 6H).
实施例6、7

Examples 6 and 7

步骤1step 1
将四氢呋喃(25mL)，环丁砜(2g,16.64mmol)加入反应瓶中，开始搅拌；温度降至-65～-70℃后，将正丁基锂(2.5M,6.66mL)滴加到其中后，温度升至-50℃反应1小时；然后将温度降至-65～-70℃后，将化合物4a(1.42g,8.32mmol)溶于四氢呋喃(25mL)中滴加到反应液，继续反应1小时。加入100mL饱和氯化铵溶液淬灭，分液；水相用乙酸乙酯(25mL)萃取，合并有机相，饱和食盐水(50mL)洗涤一次后，得到的有机相加入无水硫酸钠干燥后，过滤，滤液在45℃减压浓缩得到粗品，再加入10mL甲基叔丁醚混合搅拌0.5小时后，过滤，得到化合物6b。MS m/z:192.1[M-100+1]+；1H NMR(400MHz,CDCl3)δ4.20(d,J＝9.6Hz,1H),4.04(d,J＝9.2Hz,1H),3.95(d,J＝9.6Hz,1H),3.81(d,J＝9.2Hz,1H),3.68(s,1H),3.48(d,J＝8.2Hz,1H),3.22-3.15(m,1H),3.03-2.94(m,1H),2.37-2.08(m,4H),1.44(s,9H)。Add tetrahydrofuran (25mL) and sulfolane (2g, 16.64mmol) into the reaction flask and start stirring; after the temperature drops to -65~-70°C, n-butyllithium (2.5M, 6.66mL) is added dropwise into it, The temperature was raised to -50°C and the reaction was carried out for 1 hour; then, after the temperature was lowered to -65~-70°C, compound 4a (1.42g, 8.32mmol) was dissolved in tetrahydrofuran (25mL) and added dropwise to the reaction solution, and the reaction was continued for 1 hour. . Add 100 mL of saturated ammonium chloride solution to quench and separate the layers; extract the aqueous phase with ethyl acetate (25 mL), combine the organic phases, wash once with saturated brine (50 mL), and dry the obtained organic phase by adding anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure at 45°C to obtain a crude product. Then 10 mL of methyl tert-butyl ether is added, mixed and stirred for 0.5 hours, and then filtered to obtain compound 6b. MS m/z: 192.1[M-100+1] + ; 1 H NMR (400MHz, CDCl 3 ) δ4.20 (d, J = 9.6 Hz, 1H), 4.04 (d, J = 9.2 Hz, 1H), 3.95(d,J＝9.6Hz,1H),3.81(d,J＝9.2Hz,1H),3.68(s,1H),3.48(d,J＝8.2Hz,1H),3.22-3.15(m,1H ),3.03-2.94(m,1H),2.37-2.08(m,4H),1.44(s,9H).
步骤2Step 2
将二氯甲烷(7.5mL)，化合物6b(0.5g,1.72mmol)加入反应瓶中，开始搅拌；然后将温度降至-20℃，将二乙氨基三氟化硫(290.44mg,1.80mmol)滴加到其中，反应2小时。向反应体系中加入10mL水和10mL二氯甲烷，分液；有机相依次用饱和碳酸氢钠溶液(10mL)，饱和食盐水洗涤(10mL)后，加入无水硫酸钠干燥后，过滤，滤液在45℃下减压浓缩得到粗品，再经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～10:90)，得到粗品。再与2mL甲基叔丁醚混合搅拌，过滤，滤液在45℃下进行减压浓缩。得到化合物6c与化合物7c的混合物。化合物6c：MS m/z:174.0[M-100+1]+；化合物7c：MS m/z:194.0[M-100+1]+。Add dichloromethane (7.5 mL) and compound 6b (0.5 g, 1.72 mmol) into the reaction flask and start stirring; then lower the temperature to -20°C, and add diethylamino sulfur trifluoride (290.44 mg, 1.80 mmol) Add it dropwise and react for 2 hours. Add 10 mL of water and 10 mL of methylene chloride to the reaction system, and separate the layers; the organic phase was washed with saturated sodium bicarbonate solution (10 mL) and saturated brine (10 mL), and then dried with anhydrous sodium sulfate, filtered, and the filtrate was Concentrate under reduced pressure at 45°C to obtain a crude product, which is then separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:0 to 10:90) to obtain a crude product. Then mix and stir with 2 mL of methyl tert-butyl ether, filter, and the filtrate is concentrated under reduced pressure at 45°C. A mixture of compound 6c and compound 7c was obtained. Compound 6c: MS m/z: 174.0[M-100+1] + ; Compound 7c: MS m/z: 194.0[M-100+1] + .
步骤3Step 3
将乙酸乙酯(1mL)，化合物6c与7c的混合物(0.1g,365.83μmol)加入反应瓶中，开始搅拌；然后将氯化氢/乙酸乙酯(4M,2mL)加入其中，室温18℃，反应2小时。反应液直接过滤，45℃下进行减压浓缩得到化合物6d的盐酸盐和7d的盐酸盐混合物。Add ethyl acetate (1 mL) and the mixture of compounds 6c and 7c (0.1 g, 365.83 μmol) into the reaction flask and start stirring; then add hydrogen chloride/ethyl acetate (4 M, 2 mL) at room temperature 18°C, reaction 2 Hour. The reaction solution was directly filtered and concentrated under reduced pressure at 45°C to obtain a mixture of the hydrochloride salt of compound 6d and the hydrochloride salt of 7d.
步骤4Step 4
将异丙醇(3mL)，化合物4h(60mg,248.84μmol)加入反应瓶中。开始搅拌，然后将化合物6d的盐酸盐和7d的盐酸盐混合物(56.04mg,267.25μmol)，三乙胺(125.90mg,1.24mmol,173.18μL)加入其中，升温至100℃，反应12小时。向反应体系中加入15mL饱和氯化铵溶液以及15mL乙酸乙酯后，分液；水相用乙酸乙酯乙酯(15mL)洗涤后，合并有机相，用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃减压浓缩。粗品经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝1:4)，得到化合物6e和化合物7e的混合物。化合物6e：MS m/z:378.0[M+1]+；化合物7e：MS m/z:398.0[M+1]+。 Isopropyl alcohol (3 mL) and compound 4h (60 mg, 248.84 μmol) were added to the reaction bottle. Start stirring, then add the mixture of the hydrochloride salt of compound 6d and the hydrochloride salt of 7d (56.04 mg, 267.25 μmol) and triethylamine (125.90 mg, 1.24 mmol, 173.18 μL), and raise the temperature to 100°C and react for 12 hours. . Add 15 mL of saturated ammonium chloride solution and 15 mL of ethyl acetate to the reaction system, and separate the layers; wash the aqueous phase with ethyl acetate (15 mL), combine the organic phases, wash with 15 mL of saturated brine, and add anhydrous After drying over sodium sulfate, it was filtered, and the filtrate was concentrated under reduced pressure at 45°C. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:4) to obtain a mixture of compound 6e and compound 7e. Compound 6e: MS m/z: 378.0[M+1] + ; Compound 7e: MS m/z: 398.0[M+1] + .
步骤5Step 5
将1,4-二氧六环(1.5mL)，化合物6e和7e混合物(30mg,79.39μmol)加入反应瓶中，开始搅拌；然后将碳酸铯(77.60mg,238.17μmol)，化合物1h-2(17.96mg,79.39μmol)加入其中，氮气置换后将BrettPhos Pd G3(21.59mg,23.82μmol)加入其中，升温至100℃，反应12小时。向反应体系中加入15mL饱和氯化铵溶液以及15mL乙酸乙酯；得到的水相用乙酸乙酯(15mL×2)萃取后，合并有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩。粗品经薄层色谱法纯化(乙酸乙酯：Add 1,4-dioxane (1.5mL), the mixture of compounds 6e and 7e (30mg, 79.39μmol) into the reaction bottle and start stirring; then add cesium carbonate (77.60mg, 238.17μmol), compound 1h-2 ( 17.96 mg, 79.39 μmol) was added, and after nitrogen replacement, BrettPhos Pd G3 (21.59 mg, 23.82 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 12 hours. Add 15 mL saturated ammonium chloride solution and 15 mL ethyl acetate to the reaction system; extract the obtained aqueous phase with ethyl acetate (15 mL × 2), wash the combined organic phases with 15 mL saturated brine, and add anhydrous sodium sulfate to dry it. Afterwards, it was filtered, and the filtrate was concentrated under reduced pressure at 45°C. The crude product was purified by thin layer chromatography (ethyl acetate:
甲醇＝15:1)，得到化合物6粗品和化合物7粗品。Methanol = 15:1) to obtain crude compound 6 and crude compound 7.
化合物6粗品用1.5mL甲基叔丁基醚/二氯甲烷(5:1)混合溶液搅拌15分钟后，过滤，得到化合物6。MS m/z:568.3[M+1]+；1H NMR(400MHz,CD3OD)δ9.08(s,1H),8.56(s,1H),8.00(d,J＝6.0Hz,1H),7.95(s,1H),6.42(d,J＝5.2Hz,1H),5.29(s,2H),5.12(s,2H),4.75-4.68(m,2H),3.73-3.64(m,1H),3.51-3.43(m,1H),3.28-3.24(m,2H),3.16-3.14(m,2H),2.79-2.74(m,2H),2.28-2.21(m,2H),1.96-1.85(m,2H),1.49-1.38(m,9H)。The crude compound 6 was stirred with 1.5 mL of methyl tert-butyl ether/dichloromethane (5:1) mixed solution for 15 minutes, and then filtered to obtain compound 6. MS m/z:568.3[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δ9.08 (s, 1H), 8.56 (s, 1H), 8.00 (d, J = 6.0Hz, 1H) ,7.95(s,1H),6.42(d,J＝5.2Hz,1H),5.29(s,2H),5.12(s,2H),4.75-4.68(m,2H),3.73-3.64(m,1H ),3.51-3.43(m,1H),3.28-3.24(m,2H),3.16-3.14(m,2H),2.79-2.74(m,2H),2.28-2.21(m,2H),1.96-1.85 (m,2H),1.49-1.38(m,9H).
化合物7粗品经制备高效液相色谱分离(色谱柱:Phenomenex C18 75×30mm×3μm；流动相:A(乙腈)和B(水，含0.05％氨水和0.01mol碳酸氢铵)；梯度：B％:5％-45％)，得到化合物7。MS m/z:588.3[M+1]+；1H NMR(400MHz,CD3OD)δ9.09(s,1H),8.57(s,1H),8.00(d,J＝6.0Hz,1H),7.93(s,1H),6.43(d,J＝5.6Hz,1H),4.80-4.59(m,6H),3.88-3.78(m,1H),3.73-3.64(m,1H),3.50-3.42(m,1H),3.26-3.18(m,3H),3.13-3.06(m,1H),2.61-2.51(m,1H),2.36-2.13(m,3H),1.95-1.85(m,2H),1.50-1.35(m,9H)。The crude compound 7 was separated by preparative high performance liquid chromatography (chromatographic column: Phenomenex C18 75×30mm×3μm; mobile phase: A (acetonitrile) and B (water, containing 0.05% ammonia water and 0.01mol ammonium bicarbonate); gradient: B% :5%-45%) to obtain compound 7. MS m/z:588.3[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δ9.09 (s, 1H), 8.57 (s, 1H), 8.00 (d, J = 6.0Hz, 1H) ,7.93(s,1H),6.43(d,J＝5.6Hz,1H),4.80-4.59(m,6H),3.88-3.78(m,1H),3.73-3.64(m,1H),3.50-3.42 (m,1H),3.26-3.18(m,3H),3.13-3.06(m,1H),2.61-2.51(m,1H),2.36-2.13(m,3H),1.95-1.85(m,2H) ,1.50-1.35(m,9H).
实施例8
Example 8
步骤1step 1
将乙酸乙酯(5mL)，化合物6b(1g,3.43mmol)加入反应瓶中，开始搅拌；然后将氯化氢/乙酸乙酯(4M,15mL)加入其中，室温18℃，反应2小时。反应液直接过滤，滤饼在45℃下真空干燥得到化合物8a的盐酸盐。Add ethyl acetate (5 mL) and compound 6b (1 g, 3.43 mmol) into the reaction flask and start stirring; then add hydrogen chloride/ethyl acetate (4 M, 15 mL) and react at room temperature 18°C for 2 hours. The reaction solution was directly filtered, and the filter cake was dried under vacuum at 45°C to obtain the hydrochloride of compound 8a.
步骤2Step 2
将异丙醇(2mL)，化合物4h(80mg,331.79μmol)加入反应瓶中。开始搅拌，然后将化合物8a的盐酸盐(95.18mg,497.69μmol)，三乙胺(134.30mg,1.33mmol,184.73μL)加入其中，升温至100℃，反应2小时。向反应液中加入15mL饱和氯化铵溶液以及15mL乙酸乙酯萃取；得到的水相用15mL乙酸乙酯萃取后，合并有机相，用饱和食盐水15mL洗涤后，加入无水硫酸钠干燥，过滤，滤液在50℃下进行减压浓缩得粗品。粗品再经硅胶柱层析分离纯化(石油醚:乙酸乙＝5:1～1:4)，得到化合物8b。MS m/z:396.0[M+1]+； 1H NMR(400MHz,CD3OD)δ＝9.11(s,1H),8.06(s,1H),7.88(s,1H),4.69(dd,J＝21.6Hz,10.4Hz,2H),4.41(dd,J＝19.6Hz,10.0Hz,2H),3.62-3.58(m,1H),3.40-3.35(m,1H),3.15-3.05(m,2H),2.51-2.44(m,1H),2.31-2.20(m,2H),2.16-2.07(m,1H),1.35(d,J＝6.8Hz,6H)。Isopropyl alcohol (2 mL) and compound 4h (80 mg, 331.79 μmol) were added to the reaction bottle. Start stirring, then add the hydrochloride of compound 8a (95.18 mg, 497.69 μmol) and triethylamine (134.30 mg, 1.33 mmol, 184.73 μL), raise the temperature to 100°C, and react for 2 hours. Add 15 mL of saturated ammonium chloride solution and 15 mL of ethyl acetate to the reaction solution for extraction; extract the obtained aqueous phase with 15 mL of ethyl acetate, combine the organic phases, wash with 15 mL of saturated brine, add anhydrous sodium sulfate, dry, and filter , the filtrate was concentrated under reduced pressure at 50°C to obtain crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1~1:4) to obtain compound 8b. MS m/z:396.0[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δ = 9.11 (s, 1H), 8.06 (s, 1H), 7.88 (s, 1H), 4.69 (dd, J = 21.6Hz, 10.4Hz, 2H), 4.41 ( dd,J＝19.6Hz,10.0Hz,2H),3.62-3.58(m,1H),3.40-3.35(m,1H),3.15-3.05(m,2H),2.51-2.44(m,1H),2.31 -2.20(m,2H),2.16-2.07(m,1H),1.35(d,J＝6.8Hz,6H).
步骤3Step 3
将1,4-二氧六环(2mL)，化合物8b(40mg,101.03μmol)加入反应瓶中，开始搅拌；然后将碳酸铯(98.76mg,303.10μmol)，化合物1h-2(20.57mg,90.93μmol)加入其中，氮气置换后将BrettPhos Pd G3(27.48mg,30.31μmol)加入其中，升温至100℃，反应5小时。向反应体系中加入15mL饱和氯化铵溶液以及15mL乙酸乙酯；得到的水相用乙酸乙酯(15mL×2)萃取后，合并有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层层析法(乙酸乙酯)纯化，得到化合物8。MS m/z:586.3[M+1]+；1H NMR(400MHz,CD3OD)δ9.08(s,1H),8.52(s,1H),7.99(d,J＝5.6Hz,1H),7.89(s,1H),6.41(d,J＝6.0Hz,1H),4.74-4.61(m,4H),4.39(dd,J＝20.0and 9.6Hz,2H),3.73-3.59(m,2H),3.45-3.38(m,1H),3.27-3.20(m,2H),3.17-3.04(m,2H),2.52-2.45(m,1H),2.32-2.22(m,2H),2.17-2.07(m,1H),1.94-1.84(m,2H),1.48-1.37(m,9H)。Add 1,4-dioxane (2mL), compound 8b (40mg, 101.03μmol) into the reaction bottle and start stirring; then add cesium carbonate (98.76mg, 303.10μmol), compound 1h-2 (20.57mg, 90.93 μmol) was added, and after nitrogen replacement, BrettPhos Pd G3 (27.48 mg, 30.31 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 5 hours. Add 15 mL saturated ammonium chloride solution and 15 mL ethyl acetate to the reaction system; extract the obtained aqueous phase with ethyl acetate (15 mL × 2), wash the combined organic phases with 15 mL saturated brine, and add anhydrous sodium sulfate to dry it. Afterwards, it was filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was purified by thin layer chromatography (ethyl acetate) to obtain compound 8. MS m/z:586.3[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δ9.08 (s, 1H), 8.52 (s, 1H), 7.99 (d, J = 5.6Hz, 1H) ,7.89(s,1H),6.41(d,J＝6.0Hz,1H),4.74-4.61(m,4H),4.39(dd,J＝20.0and 9.6Hz,2H),3.73-3.59(m,2H ),3.45-3.38(m,1H),3.27-3.20(m,2H),3.17-3.04(m,2H),2.52-2.45(m,1H),2.32-2.22(m,2H),2.17-2.07 (m,1H),1.94-1.84(m,2H),1.48-1.37(m,9H).
实施例9
Example 9
步骤1step 1
将湿钯碳(50mg，548.75μmol，10％含量)加入反应瓶中，加入乙醇(5mL)，再加入化合物6c和7c的混合物(150mg，548.75μmol)氢气置换三次，氢气，25℃，15psi下搅拌3小时。反应完成后将反应液过滤后浓缩得到化合物9a。MS m/z:220.0[M-56+1]+；1H NMR(400MHz,CDCl3)δppm 4.08-4.23(m,2H),3.85-3.97(m,1H),3.63-3.77(m,1H),3.23-3.34(m,1H),3.09-3.21(m,1H),2.96-3.08(m,1H),2.77-2.93(m,1H),2.33-2.50(m,1H),2.18-2.31(m,1H),2.02-2.16(m,1H),1.75-1.85(m,1H),1.36-1.52(m,9H)。Add wet palladium on carbon (50 mg, 548.75 μmol, 10% content) into the reaction bottle, add ethanol (5 mL), and then add the mixture of compounds 6c and 7c (150 mg, 548.75 μmol) and replace it with hydrogen three times, hydrogen, 25°C, 15 psi Stir for 3 hours. After the reaction is completed, the reaction solution is filtered and concentrated to obtain compound 9a. MS m/z:220.0[M-56+1] + ; 1 H NMR (400MHz, CDCl 3 ) δppm 4.08-4.23(m,2H),3.85-3.97(m,1H),3.63-3.77(m,1H ),3.23-3.34(m,1H),3.09-3.21(m,1H),2.96-3.08(m,1H),2.77-2.93(m,1H),2.33-2.50(m,1H),2.18-2.31 (m,1H),2.02-2.16(m,1H),1.75-1.85(m,1H),1.36-1.52(m,9H).
步骤2Step 2
将化合物9a(140mg，508.42μmol)溶于乙酸乙酯(0.5mL)，加入氯化氢/乙酸乙酯(3mL)，20℃下搅拌4小时。反应完成后将反应液直接浓缩得化合物9b的盐酸盐。MS m/z:176.1[M+1]+；1H NMR(400MHz,CD3OD)δ＝4.21(q,J＝10.0Hz,2H),4.15-4.02(m,2H),3.61(q,J＝7.0Hz,1H),3.53-.44(m,1H),3.22-3.11(m,1H),3.09-2.98(m,1H),2.47-2.36(m,1H),2.26-2.06(m,2H),2.02(d,J＝3.5Hz,1H),1.88-1.74(m,1H)。Compound 9a (140 mg, 508.42 μmol) was dissolved in ethyl acetate (0.5 mL), hydrogen chloride/ethyl acetate (3 mL) was added, and the mixture was stirred at 20°C for 4 hours. After the reaction is completed, the reaction solution is directly concentrated to obtain the hydrochloride of compound 9b. MS m/z: 176.1[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δ = 4.21 (q, J = 10.0Hz, 2H), 4.15-4.02 (m, 2H), 3.61 (q, J＝7.0Hz,1H),3.53-.44(m,1H),3.22-3.11(m,1H),3.09-2.98(m,1H),2.47-2.36(m,1H),2.26-2.06(m ,2H),2.02(d,J＝3.5Hz,1H),1.88-1.74(m,1H).
步骤3Step 3
在反应瓶中加入异丙醇(2mL)，化合物4h(90mg,373.26μmol)，化合物9b的盐酸盐(94.83mg,447.92μmol)，三乙胺(113.31mg,1.12mmol,155.86μL)氮气置换三次，100℃搅拌3小时。反应完成后将反应液 进行减压浓缩得到粗品。粗品通过柱层析纯化(石油醚：乙酸乙酯＝10:90)得到化合物9c，在此纯化过程除去化合物7c衍生的化合物7e。MS m/z:380.0[M+1]+；1H NMR(400MHz,CD3OD)δppm 9.07-9.16(m,1H),8.04(s,1H),7.86(s,1H),4.67(dt,J＝11.04,8.80Hz,2H),4.42(dd,J＝9.01,5.63Hz,1H),4.34(dd,J＝8.82,5.69Hz,1H),3.48-3.61(m,1H),3.34-3.41(m,1H),3.15-3.26(m,2H),3.02-3.12(m,1H),2.43-2.56(m,1H),2.20-2.30(m,1H),2.07-2.17(m,1H),1.83-1.96(m,1H),1.32-1.40(m,6H)。Add isopropanol (2mL), compound 4h (90mg, 373.26μmol), compound 9b hydrochloride (94.83mg, 447.92μmol), triethylamine (113.31mg, 1.12mmol, 155.86μL) to the reaction bottle with nitrogen replacement. Three times, stir at 100°C for 3 hours. After the reaction is completed, the reaction solution Concentrate under reduced pressure to obtain crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:90) to obtain compound 9c. During this purification process, compound 7e derived from compound 7c was removed. MS m/z:380.0[M+1] + ; 1 H NMR (400MHz, CD 3 OD) δppm 9.07-9.16(m,1H),8.04(s,1H),7.86(s,1H),4.67(dt ,J＝11.04,8.80Hz,2H),4.42(dd,J＝9.01,5.63Hz,1H),4.34(dd,J＝8.82,5.69Hz,1H),3.48-3.61(m,1H),3.34- 3.41(m,1H),3.15-3.26(m,2H),3.02-3.12(m,1H),2.43-2.56(m,1H),2.20-2.30(m,1H),2.07-2.17(m,1H ),1.83-1.96(m,1H),1.32-1.40(m,6H).
步骤4Step 4
在反应瓶中加入化合物9c(50mg,131.61μmol)，化合物1h-2(32.76mg,144.77μmol)，二氧六环(5mL)，碳酸铯(85.76mg,263.22μmol)，氮气置换三次加入BrettPhos Pd G3(23.86mg,26.32μmol)，100℃搅拌5小时。反应液冷却至室温后加入水5mL，用乙酸乙酯萃取(10mL×3)，收集有机相，有机相用饱和食盐水洗涤(10mL×2)，无水硫酸钠干燥，过滤，减压浓缩得到产物粗品。粗品通过硅胶柱层析纯化(二氯甲烷：甲醇＝100:0-10:1)得到产物粗品。得到的产物粗品中加入乙酸乙酯0.5mL，正庚烷1.5mL搅拌15min后过滤，收集滤饼，滤饼减压浓缩得到化合物9。MS m/z:570.0[M+1]+；1H NMR(400MHz,CDCl3)δppm 8.97-9.05(m,1H),8.39-8.46(m,1H),8.08-8.15(m,1H),8.00-8.08(m,1H),7.37-7.45(m,1H),6.17-6.24(m,1H),4.71-4.80(m,1H),4.54-4.65(m,2H),4.41-4.51(m,2H),4.15-4.24(m,1H),3.68-3.81(m,1H),3.49-3.64(m,2H),3.34-3.49(m,2H),3.14-.27(m,2H),2.99-3.11(m,1H),2.42-2.54(m,1H),2.23-2.34(m,1H),2.11-2.22(m,1H),1.85-2.07(m,4H),1.46-1.56(m,3H),1.34-1.41(m,6H)。Add compound 9c (50 mg, 131.61 μmol), compound 1h-2 (32.76 mg, 144.77 μmol), dioxane (5 mL), and cesium carbonate (85.76 mg, 263.22 μmol) into the reaction bottle. Replace with nitrogen three times and add BrettPhos Pd. G3 (23.86 mg, 26.32 μmol), stirred at 100°C for 5 hours. After the reaction solution was cooled to room temperature, 5 mL of water was added, extracted with ethyl acetate (10 mL × 3), the organic phase was collected, washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain Crude product. The crude product was purified by silica gel column chromatography (dichloromethane: methanol = 100:0-10:1) to obtain the crude product. To the obtained crude product, 0.5 mL of ethyl acetate and 1.5 mL of n-heptane were added, stirred for 15 min, filtered, and the filter cake was collected, and the filter cake was concentrated under reduced pressure to obtain compound 9. MS m/z:570.0[M+1] + ; 1 H NMR (400MHz, CDCl 3 ) δppm 8.97-9.05(m,1H),8.39-8.46(m,1H),8.08-8.15(m,1H), 8.00-8.08(m,1H),7.37-7.45(m,1H),6.17-6.24(m,1H),4.71-4.80(m,1H),4.54-4.65(m,2H),4.41-4.51(m ,2H),4.15-4.24(m,1H),3.68-3.81(m,1H),3.49-3.64(m,2H),3.34-3.49(m,2H),3.14-.27(m,2H), 2.99-3.11(m,1H),2.42-2.54(m,1H),2.23-2.34(m,1H),2.11-2.22(m,1H),1.85-2.07(m,4H),1.46-1.56(m ,3H),1.34-1.41(m,6H).
实施例10
Example 10
步骤1step 1
将化合物10a(5g,24.27mmol)，异丙胺(4.30g,72.81mmol,6.26mL)溶于EtOH(20mL)，80℃搅拌反应16小时。将反应液减压浓缩，柱层析(石油醚:乙酸乙酯＝20:1)纯化，得到化合物10b和10b'的混合物，含量接近1:1。MS m/z:228.9[M+1]+。MS m/z:242.9[M+1]+。Compound 10a (5g, 24.27mmol) and isopropylamine (4.30g, 72.81mmol, 6.26mL) were dissolved in EtOH (20mL), and the reaction was stirred at 80°C for 16 hours. The reaction solution was concentrated under reduced pressure and purified by column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain a mixture of compounds 10b and 10b', with a content close to 1:1. MS m/z:228.9[M+1] + . MS m/z:242.9[M+1] + .
步骤2 Step 2
将化合物10b和10b'的混合物(5g,21.87mmol)溶解在THF(50mL)\MeOH(5mL)，H2O(50mL)中，加入一水合氢氧化锂(4.59g,109.33mmol)，20℃下搅拌4小时。加稀盐酸调节pH至6，乙酸乙酯(60mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤减压浓缩得到粗品化合物10c，直接用于下一步。MS m/z:214.8[M+1]+。Dissolve the mixture of compounds 10b and 10b' (5g, 21.87mmol) in THF (50mL)\MeOH (5mL), H 2 O (50mL), add lithium hydroxide monohydrate (4.59g, 109.33mmol), 20°C Stir for 4 hours. Add dilute hydrochloric acid to adjust the pH to 6, extract with ethyl acetate (60 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure to obtain crude compound 10c, which is used directly in the next step. MS m/z:214.8[M+1] + .
步骤3Step 3
将化合物10c(4.5g,20.96mmol)和HOBt(2.83g,20.96mmol)溶解在THF(90mL)中，0℃下滴加DCC(4.33g,20.96mmol,4.24mL)溶解在THF(30mL)中，搅拌6小时，过滤，得到化合物10d的四氢呋喃溶液直接用于下一步。将丙二酸二乙酯(3.36g,20.96mmol,3.17mL)溶解在THF(30mL)中，0℃下加入NaH(3.35g,83.84mmol,60％)搅拌30分钟，再加入最开始反应得到的化合物10d四氢呋喃溶液，20℃继续搅拌11.5小时。将反应液加稀盐酸淬灭，pH至5，乙酸乙酯萃取(60mL×2)，合并有机相，无水硫酸钠干燥，过滤浓缩得到化合物10e。MS m/z:310.9[M+1]+。Compound 10c (4.5g, 20.96mmol) and HOBt (2.83g, 20.96mmol) were dissolved in THF (90mL), and DCC (4.33g, 20.96mmol, 4.24mL) was added dropwise at 0°C to dissolve it in THF (30mL). , stir for 6 hours, filter, and obtain a tetrahydrofuran solution of compound 10d, which is directly used in the next step. Dissolve diethyl malonate (3.36g, 20.96mmol, 3.17mL) in THF (30mL), add NaH (3.35g, 83.84mmol, 60%) at 0°C and stir for 30 minutes, then add the initial reaction to obtain A solution of compound 10d in tetrahydrofuran was stirred at 20°C for 11.5 hours. The reaction solution was quenched by adding dilute hydrochloric acid to pH 5, and extracted with ethyl acetate (60 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 10e. MS m/z:310.9[M+1] + .
步骤4Step 4
将化合物10e(6g,19.31mmol,粗品)加入到三氟乙酸(15.40g,135.06mmol,10mL)和浓盐酸(10.20g,103.51mmol,10mL,37％含量)的混合体系中，加热到80℃，反应13小时。将反应液减压浓缩，再用2M NaOH慢慢中和至中性，乙酸乙酯(100mL×2)萃取，合并有机相，水洗，无水硫酸钠干燥，过滤浓缩得到粗品化合物10f直接用于下一步。MS m/z:238.8[M+1]+。Compound 10e (6g, 19.31mmol, crude product) was added to a mixed system of trifluoroacetic acid (15.40g, 135.06mmol, 10mL) and concentrated hydrochloric acid (10.20g, 103.51mmol, 10mL, 37% content), and heated to 80°C. , react for 13 hours. The reaction solution was concentrated under reduced pressure, then slowly neutralized with 2M NaOH until neutral, extracted with ethyl acetate (100mL×2), combined the organic phases, washed with water, dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude compound 10f, which was directly used Next step. MS m/z:238.8[M+1] + .
步骤5Step 5
将化合物10f(2.6g,10.89mmol)加入到DCM(30mL)中，加入三乙胺(4.41g,43.57mmol,6.07mL)0℃下再滴加三氟甲磺酸酐(9.22g,32.68mmol,5.39mL)反应2小时。加水(50mL)淬灭，分液，水相用二氯甲烷(50mL)再萃取一次，合并有机相，无水硫酸钠干燥，过滤浓缩得粗品。粗品经色谱柱纯化(石油醚:乙酸乙酯＝5:1)得到产品得到化合物10g。MS m/z:370.7[M+1]+。1H NMR(400MHz,CDCl3)δ8.77(s,1H),7.47(s,1H),6.76(s,1H),5.22-5.30(m,1H),1.66(d,J＝4.8Hz,6H)。Add compound 10f (2.6g, 10.89mmol) to DCM (30mL), add triethylamine (4.41g, 43.57mmol, 6.07mL) and then add trifluoromethanesulfonic anhydride (9.22g, 32.68mmol) dropwise at 0°C. 5.39mL) react for 2 hours. Add water (50 mL) to quench, separate the layers, extract the aqueous phase again with dichloromethane (50 mL), combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain the product, and 10 g of the compound was obtained. MS m/z:370.7[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ8.77(s,1H),7.47(s,1H),6.76(s,1H),5.22-5.30(m,1H),1.66(d,J＝4.8Hz, 6H).
步骤6Step 6
将化合物10g(200mg,539.47μmol)、化合物1c(213.01mg,809.21μmol,三氟乙酸盐)、碳酸铯(527.31mg,1.62mmol)、Xantphos Pd G4(51.92mg,53.95μmol)分别加入到1,4-二氧六环(5mL)中，氮气置换，升温至100℃，搅拌4小时。反应液冷却，加水20mL，乙酸乙酯萃取(30mL×2)，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩得粗品。粗品经柱层析纯化(DCM:MeOH＝20:1～10:1)得到产物化合物10h。MS m/z:369.8[M+1]+。Compound 10g (200mg, 539.47μmol), compound 1c (213.01mg, 809.21μmol, trifluoroacetate), cesium carbonate (527.31mg, 1.62mmol), and Xantphos Pd G4 (51.92mg, 53.95μmol) were added to 1 , 4-dioxane (5 mL) was replaced with nitrogen, the temperature was raised to 100°C, and stirred for 4 hours. The reaction solution was cooled, 20 mL of water was added, extracted with ethyl acetate (30 mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (DCM:MeOH=20:1～10:1) to obtain the product compound 10h. MS m/z:369.8[M+1] + .
步骤7Step 7
将化合物10h(20mg,54.07μmol)，化合物1h-2(24.47mg,108.15μmol)加入到1,4-二氧六环(1mL)中，加入Cs2CO3(52.85mg,162.22μmol)，BrettPhos Pd G3(4.90mg,5.41μmol)，Xantphos Pd G4(5.20mg,5.41μmol)，加热升温至100℃，搅拌16小时。将反应液过滤，浓缩得粗品。再用乙腈溶解，过滤，滤液送制备色谱柱分离(色谱柱:Welch Xtimate C18 100×40mm×3μm；流动相:[水(三氟乙酸)-乙腈]；ACN％:0％-40％,8min)得到化合物10。MS m/z:560.0[M+1]+。Compound 10h (20 mg, 54.07 μmol) and compound 1h-2 (24.47 mg, 108.15 μmol) were added to 1,4-dioxane (1 mL), and Cs 2 CO 3 (52.85 mg, 162.22 μmol) was added. BrettPhos Pd G3 (4.90 mg, 5.41 μmol), Xantphos Pd G4 (5.20 mg, 5.41 μmol), heat to 100°C, and stir for 16 hours. The reaction solution was filtered and concentrated to obtain crude product. Then dissolve with acetonitrile, filter, and send the filtrate to a preparative chromatography column for separation (chromatographic column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; ACN%: 0%-40%, 8min ) to obtain compound 10. MS m/z:560.0[M+1] + .
实施例11、12
Examples 11 and 12
步骤1step 1
将二氯甲烷(6mL)，化合物4i(85mg,229.81μmol)加入反应瓶中，开始搅拌；然后将温度降至0℃后，将二乙氨基三氟化硫(38.90mg,241.30μmol)滴加到其中，反应10分钟。向反应体系中加入15mL水和15mL二氯甲烷后，分液；有机相依次用饱和碳酸氢钠溶液(15mL)，饱和食盐水(15mL)洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃减压浓缩得到粗品，再经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝1:1～1:4)得到化合物11a和12a。化合物11a MS m/z:372.0[M+1]+。1H NMR(400MHz,CD3OD)δppm9.10(s,1H),8.10(s,1H),7.91(s,1H),4.85-4.77(m,2H),4.72-4.64(m,2H),4.02(d,J＝22.4Hz,2H),3.43-3.36(m,1H),3.07(s,3H),1.35(d,J＝6.8Hz,6H).化合物12a MS m/z:352.0[M+1]+。Add dichloromethane (6 mL) and compound 4i (85 mg, 229.81 μmol) into the reaction flask and start stirring; then, after the temperature drops to 0°C, diethylamino sulfur trifluoride (38.90 mg, 241.30 μmol) is added dropwise. into it and react for 10 minutes. After adding 15 mL of water and 15 mL of dichloromethane to the reaction system, separate the liquids; wash the organic phase with saturated sodium bicarbonate solution (15 mL) and saturated brine (15 mL), add anhydrous sodium sulfate, dry, filter, and the filtrate Concentrate under reduced pressure at 45°C to obtain a crude product, which is then separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1 to 1:4) to obtain compounds 11a and 12a. Compound 11a MS m/z: 372.0[M+1] + . 1 H NMR (400MHz, CD 3 OD) δppm9.10(s,1H),8.10(s,1H),7.91(s,1H),4.85-4.77(m,2H),4.72-4.64(m,2H) ,4.02(d,J＝22.4Hz,2H),3.43-3.36(m,1H),3.07(s,3H),1.35(d,J＝6.8Hz,6H). Compound 12a MS m/z:352.0[ M+1] + .
步骤2Step 2
将1,4-二氧六环(4mL)，化合物11a(25mg,67.23μmol)加入反应瓶中，开始搅拌；然后将乙酸钾(19.79mg,201.69μmol)，化合物1h-2(22.82mg,100.85μmol)加入其中，氮气置换后将BrettPhos Pd G3(18.28mg,20.17μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入10mL饱和氯化铵溶液以及10mL乙酸乙酯；得到的水相用乙酸乙酯(10mL×2)萃取后，合并有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得粗品，再经薄层层析法(石油醚：乙酸乙酯＝1:4)纯化，得到化合物11。MS m/z:562.3[M+1]+。1H NMR(400MHz,CD3OD)δppm 9.07(s,1H),8.56(s,1H),8.00(d,J＝5.6Hz,1H),7.93(s,1H),6.43(d,J＝5.6Hz,1H),4.83-4.61(m,6H),4.03(d,J＝22.4Hz,2H),3.73-3.64(m,1H),3.48-3.42(m,1H),3.28-3.20(m,2H),3.08(s,3H),1.94-1.87(m,2H),1.46(d,J＝21.2Hz,3H),1.38(d,J＝6.8Hz,6H). Add 1,4-dioxane (4mL), compound 11a (25mg, 67.23μmol) into the reaction bottle and start stirring; then add potassium acetate (19.79mg, 201.69μmol), compound 1h-2 (22.82mg, 100.85 μmol) was added, and after nitrogen replacement, BrettPhos Pd G3 (18.28 mg, 20.17 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. Add 10 mL saturated ammonium chloride solution and 10 mL ethyl acetate to the reaction system; extract the obtained aqueous phase with ethyl acetate (10 mL × 2), wash the combined organic phases with 15 mL saturated brine, and add anhydrous sodium sulfate to dry it. , filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product, which was then purified by thin layer chromatography (petroleum ether: ethyl acetate = 1:4) to obtain compound 11. MS m/z:562.3[M+1] + . 1 H NMR (400MHz, CD 3 OD) δppm 9.07 (s, 1H), 8.56 (s, 1H), 8.00 (d, J＝5.6Hz, 1H), 7.93 (s, 1H), 6.43 (d, J＝ 5.6Hz,1H),4.83-4.61(m,6H),4.03(d,J＝22.4Hz,2H),3.73-3.64(m,1H),3.48-3.42(m,1H),3.28-3.20(m ,2H),3.08(s,3H),1.94-1.87(m,2H),1.46(d,J＝21.2Hz,3H),1.38(d,J＝6.8Hz,6H).
将1,4-二氧六环(1.5mL)，化合物12a(10mg,28.42μmol)加入反应瓶中，开始搅拌；然后将乙酸钾(8.37mg,85.26μmol)，化合物1h-2(9.65mg,42.63μmol)加入其中，氮气置换后将BrettPhos Pd G3(7.73mg,8.53μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入10mL饱和氯化铵溶液以及10mL乙酸乙酯；得到的水相用乙酸乙酯(10mL×2)萃取后，合并有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得粗品，再经薄层层析法(乙酸乙酯)纯化得到化合物12。MS m/z:542.3[M+1]+。1H NMR(400MHz,CD3OD)δppm 9.09(s,1H),8.58(s,1H),8.00(d,J＝6.0Hz,1H),7.96(s,1H),6.63(s,1H),6.43(d,J＝5.6Hz,1H),5.41-5.40(m,2H),5.17(s,2H),4.75-4.67(m,2H),3.74-3.63(m,1H),3.50-3.46(m,1H),3.26-3.21(m,2H),3.07(s,3H),1.96-1.83(m,2H),1.49-1.38(m,9H)。Add 1,4-dioxane (1.5mL), compound 12a (10mg, 28.42μmol) into the reaction bottle and start stirring; then add potassium acetate (8.37mg, 85.26μmol), compound 1h-2 (9.65mg, 42.63 μmol) was added to it, and after nitrogen replacement, BrettPhos Pd G3 (7.73 mg, 8.53 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. Add 10 mL saturated ammonium chloride solution and 10 mL ethyl acetate to the reaction system; extract the obtained aqueous phase with ethyl acetate (10 mL × 2), wash the combined organic phases with 15 mL saturated brine, and add anhydrous sodium sulfate to dry it. Afterwards, it was filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product, which was then purified by thin layer chromatography (ethyl acetate) to obtain compound 12. MS m/z:542.3[M+1] + . 1 H NMR (400MHz, CD 3 OD) δppm 9.09 (s, 1H), 8.58 (s, 1H), 8.00 (d, J = 6.0Hz, 1H), 7.96 (s, 1H), 6.63 (s, 1H) ,6.43(d,J＝5.6Hz,1H),5.41-5.40(m,2H),5.17(s,2H),4.75-4.67(m,2H),3.74-3.63(m,1H),3.50-3.46 (m,1H),3.26-3.21(m,2H),3.07(s,3H),1.96-1.83(m,2H),1.49-1.38(m,9H).
实施例13
Example 13
步骤1step 1
在反应瓶中加入化合物13a(3g,22.19mmol)，二氯甲烷(60mL)，三乙胺(5.61g,55.48mmol)降温至0℃滴加入氯甲酸苄酯(5.68g,33.29mmol)，自然升温至15℃搅拌2小时。反应液进行减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:1)得到化合物13b。1H NMR(400MHz,DMSO-d6)δppm 7.23-7.47(m,5H),5.05-5.14(m,2H),3.76-3.87(m,4H),3.12-3.22(m,4H)。Add compound 13a (3g, 22.19mmol), dichloromethane (60mL), and triethylamine (5.61g, 55.48mmol) to the reaction bottle. Cool to 0°C and add benzyl chloroformate (5.68g, 33.29mmol) dropwise. Naturally The temperature was raised to 15°C and stirred for 2 hours. The reaction solution was concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:1) to obtain compound 13b. 1 H NMR (400MHz, DMSO-d 6 ) δppm 7.23-7.47(m,5H), 5.05-5.14(m,2H), 3.76-3.87(m,4H), 3.12-3.22(m,4H).
步骤2Step 2
在反应瓶中加入化合物13b(3.5g,13.00mmol)，四氢呋喃(50mL)，氮气置换三次降温至-65℃加入二(三甲基硅)氨基锂(1M,25.99mL)，-65℃搅拌0.5小时后加入1-Boc-3-氮杂环丁酮(2.22g,13.00mmol)，-65℃搅拌0.5小时。反应液中加入饱和氯化铵水溶液(20mL)淬灭，用乙酸乙酯萃取(40mL×3)，饱和食盐水(20mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物13c。Add compound 13b (3.5g, 13.00mmol) and tetrahydrofuran (50mL) to the reaction flask, replace with nitrogen three times, cool to -65°C, add lithium bis(trimethylsilyl)amide (1M, 25.99mL), and stir at -65°C for 0.5 After an hour, 1-Boc-3-azetidinone (2.22g, 13.00mmol) was added, and the mixture was stirred at -65°C for 0.5 hours. The reaction solution was quenched by adding saturated aqueous ammonium chloride solution (20 mL), extracted with ethyl acetate (40 mL × 3), washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 13c.
步骤3 Step 3
在反应瓶中加入化合物13c(2g,4.54mmol)，氯化氢/乙酸乙酯(20mL,4M)，25℃搅拌2小时。反应液中加入饱和碳酸氢钠水溶液(20mL)调节pH＝7-8，用乙酸乙酯萃取(40mL×3)，饱和食盐水(20mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品化合物13d的盐酸盐(20mL)，无后续纯化，直接用于下一步反应得到。Compound 13c (2g, 4.54mmol) and hydrogen chloride/ethyl acetate (20mL, 4M) were added to the reaction flask, and stirred at 25°C for 2 hours. Add saturated aqueous sodium bicarbonate solution (20 mL) to the reaction solution to adjust the pH to 7-8, extract with ethyl acetate (40 mL × 3), wash with saturated brine (20 mL × 2), dry over anhydrous sodium sulfate, filter, and reduce pressure. Concentration gave crude compound 13d hydrochloride (20 mL), which was directly used in the next reaction without subsequent purification.
步骤4Step 4
在反应瓶中加入化合物4h(120mg,497.69μmol)，化合物13d(562.67mg,盐酸盐)，异丙醇(6mL)，三乙胺(201.44mg,1.99mmol)氮气置换三次，100℃搅拌3小时。反应液进行减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物13e。MS m/z:545.3[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.96-9.09(m,1H),8.10-8.16(m,1H),7.70-7.75(m,1H),7.34-7.39(m,5H),5.11-5.25(m,3H),4.48-4.62(m,1),4.30-4.44(m,1H),4.16-4.29(m,2H),3.54-3.68(m,1H),3.21-3.38(m,3H),2.89-3.15(m,3H),1.32-1.42(m,6H)。Add compound 4h (120 mg, 497.69 μmol), compound 13d (562.67 mg, hydrochloride), isopropyl alcohol (6 mL), and triethylamine (201.44 mg, 1.99 mmol) into the reaction bottle, replace with nitrogen three times, and stir at 100°C for 3 times. Hour. The reaction solution was concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 13e. MS m/z:545.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.96-9.09(m,1H),8.10-8.16(m,1H),7.70-7.75(m,1H),7.34-7.39(m,5H),5.11-5.25( m,3H),4.48-4.62(m,1),4.30-4.44(m,1H),4.16-4.29(m,2H),3.54-3.68(m,1H),3.21-3.38(m,3H), 2.89-3.15(m,3H),1.32-1.42(m,6H).
步骤5Step 5
在反应瓶中加入化合物13e(180mg,330.25μmol)，二氯甲烷(10mL)，氮气置换三次，降温至-20℃加入二乙氨基三氟化硫(55.89mg,346.76μmol)，-20℃搅拌2小时。反应液中加入饱和碳酸氢钠水溶液(10mL)淬灭后，加入二氯甲烷萃取(10mL×3)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到产物。粗品用柱层析纯化(二氯甲烷：甲醇＝20:1-10:1)纯化得到化合物13f。MS m/z:527.1[M+1]+。Add compound 13e (180 mg, 330.25 μmol) and dichloromethane (10 mL) to the reaction flask, replace with nitrogen three times, cool to -20°C, add diethylaminosulfur trifluoride (55.89 mg, 346.76 μmol), and stir at -20°C. 2 hours. The reaction solution was quenched by adding saturated aqueous sodium bicarbonate solution (10 mL), and extracted with dichloromethane (10 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the product. The crude product was purified by column chromatography (dichloromethane: methanol = 20:1-10:1) to obtain compound 13f. MS m/z:527.1[M+1] + .
步骤6Step 6
在反应瓶中加入化合物13f(110mg,208.72μmol)，化合物1h-2(51.94mg,229.59μmol)，乙酸钾(61.45mg,626.15μmol)，BrettPhos Pd G3(37.84mg,41.74μmol)，二氧六环(2mL)，氮气置换三次，100℃搅拌3小时。反应液冷却至室温后加入水(5mL)后用乙酸乙酯萃取(10mL×3)，收集有机相，有机相用饱和食盐水洗涤(4mL×2)，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品用柱层析纯化(二氯甲烷：甲醇＝30:1-10:1)纯化得到化合物13g。MS m/z:717.3[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.77-8.96(m,1H),8.34-8.43(m,1H),7.97-8.10(m,2H),7.22-7.35(m,5H),6.10-6.21(m,1H),5.21-5.34(m,2H),5.02-5.20(m,3H),4.75-4.89(m,1H),4.47-4.57(m,1H),4.23-4.43(m,1H),4.22-4.32(m,1H),3.94-4.05(m,2H),3.61-3.74(m,1H),3.44-3.57(m,2H),3.29-3.40(m,1H),2.99-3.14(m,2H),1.83-1.96(m,3H),1.38-1.56(m,6H),1.31-1.33(m,3H)。Add compound 13f (110 mg, 208.72 μmol), compound 1h-2 (51.94 mg, 229.59 μmol), potassium acetate (61.45 mg, 626.15 μmol), BrettPhos Pd G3 (37.84 mg, 41.74 μmol), and dioxane into the reaction bottle. (2 mL), replaced with nitrogen three times, and stirred at 100°C for 3 hours. After the reaction solution was cooled to room temperature, water (5 mL) was added and extracted with ethyl acetate (10 mL × 3). The organic phase was collected, washed with saturated brine (4 mL × 2), dried over anhydrous sodium sulfate, filtered, and reduced pressure. Concentrate to obtain crude product. The crude product was purified by column chromatography (dichloromethane: methanol = 30:1-10:1) to obtain compound 13g. MS m/z:717.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.77-8.96(m,1H),8.34-8.43(m,1H),7.97-8.10(m,2H),7.22-7.35(m,5H),6.10-6.21( m,1H),5.21-5.34(m,2H),5.02-5.20(m,3H),4.75-4.89(m,1H),4.47-4.57(m,1H),4.23-4.43(m,1H), 4.22-4.32(m,1H),3.94-4.05(m,2H),3.61-3.74(m,1H),3.44-3.57(m,2H),3.29-3.40(m,1H),2.99-3.14(m ,2H),1.83-1.96(m,3H),1.38-1.56(m,6H),1.31-1.33(m,3H).
步骤7Step 7
在拇指瓶中加入化合物13g(50mg,69.75μmol)，二氧化铂(111.11mg,489.30μmol)，甲醇(1mL)，氢气置换三次，15psi，25℃搅拌7小时。反应液过滤，滤饼用甲醇50mL洗涤，滤液进行减压浓缩得到粗品，通过柱层析纯化(二氯甲烷：甲醇＝20:1-10:1)得到化合物13h。MS m/z:719.4[M+1]+。Add 13g of compound (50 mg, 69.75 μmol), platinum dioxide (111.11 mg, 489.30 μmol), and methanol (1 mL) into the thumb bottle, replace with hydrogen three times, stir at 15 psi and 25°C for 7 hours. The reaction solution was filtered, and the filter cake was washed with 50 mL of methanol. The filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (dichloromethane: methanol = 20:1-10:1) to obtain compound 13h. MS m/z:719.4[M+1] + .
步骤8Step 8
在反应瓶中加入化合物13h(30mg,41.73μmol)，二氯甲烷(1mL)，三甲基碘硅烷(16.70mg,83.47μmol)，氮气置换三次，25℃搅拌12小时。反应液进行减压浓缩得到产物粗品。粗品经柱层析纯化(二氯甲烷：甲醇＝10:1-5:1)后，再经制备高效液相色谱分离(色谱柱:Phenomenex C18 75×30mm×3μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:1-35％，8min)得到化合物13。MS m/z:585.3[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.98-9.04(m,1H),8.39-8.45(m,1H),8.09-8.15(m,1H),8.00-8.07(m,1H),7.38-7.45(m,1H),6.18-6.25(m,1H),4.71-4.81(m,1H),4.55-4.65(m,2H),4.40-4.54(m,2H),4.20-4.28(m,1H),3.68-3.81(m,1H),3.50-3.64(m,2H),3.24-3.47(m,7H),2.98-3.16(m,3H),1.90-2.05(m,3H),1.54(s,3H),1.34-1.42(m,6H)。Add compound 13h (30 mg, 41.73 μmol), dichloromethane (1 mL), and trimethylsilyl iodide (16.70 mg, 83.47 μmol) into the reaction bottle, replace with nitrogen three times, and stir at 25°C for 12 hours. The reaction solution was concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography (dichloromethane: methanol = 10:1-5:1), and then separated by preparative high performance liquid chromatography (chromatographic column: Phenomenex C18 75×30mm×3μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% ammonia bicarbonate); gradient: B%: 1-35%, 8 min) to give compound 13. MS m/z:585.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.98-9.04(m,1H),8.39-8.45(m,1H),8.09-8.15(m,1H),8.00-8.07(m,1H),7.38-7.45( m,1H),6.18-6.25(m,1H),4.71-4.81(m,1H),4.55-4.65(m,2H),4.40-4.54(m,2H),4.20-4.28(m,1H), 3.68-3.81(m,1H),3.50-3.64(m,2H),3.24-3.47(m,7H),2.98-3.16(m,3H),1.90-2.05(m,3H),1.54(s,3H ),1.34-1.42(m,6H).
实施例14
Example 14
步骤1step 1
将化合物1b(260mg，1.04mmol)溶于乙酸乙酯(5mL)，加入氯化氢/乙酸乙酯(20mL)，20℃下搅拌2小时。反应液直接浓缩得1c的盐酸盐。1H NMR(400MHz,CD3OD)δ＝4.24-4.16(m,2H),4.14-4.06(m,2H),3.90(dd,J＝4.6,8.6Hz,1H),3.64-3.58(m,2H),3.09(s,1H),3.02-2.99(m,3H)。Compound 1b (260 mg, 1.04 mmol) was dissolved in ethyl acetate (5 mL), hydrogen chloride/ethyl acetate (20 mL) was added, and the mixture was stirred at 20°C for 2 hours. The reaction solution was directly concentrated to obtain the hydrochloride salt of 1c. 1 H NMR (400MHz, CD 3 OD) δ = 4.24-4.16 (m, 2H), 4.14-4.06 (m, 2H), 3.90 (dd, J = 4.6, 8.6Hz, 1H), 3.64-3.58 (m, 2H),3.09(s,1H),3.02-2.99(m,3H).
步骤2Step 2
将化合物14a(15g,72.81mmol)溶于二甲基甲酰胺(150mL)中，然后加入碳酸钾(20.13g，145.65mmol)，氰基乙酸叔丁酯(11.31g，80.09mmol，11.45mL)，置换氮气3次，100℃搅拌1hr，补加氰基乙酸叔丁酯(4.11g，29.12mmol，4.17mL)继续反应1hr。向反应液中加入2M盐酸调pH至6～7，用饱和氯化铵水溶液洗一次，用乙酸乙酯(60mL×3)萃取，分液，合并有机相，用饱和食盐水洗两次，无水硫酸钠干燥，浓缩得粗品化合物14b。MS m/z:311.0[M+1]+。Compound 14a (15g, 72.81mmol) was dissolved in dimethylformamide (150mL), and then potassium carbonate (20.13g, 145.65mmol) and tert-butyl cyanoacetate (11.31g, 80.09mmol, 11.45mL) were added. Replace nitrogen three times, stir at 100°C for 1 hour, add tert-butyl cyanoacetate (4.11g, 29.12mmol, 4.17mL) and continue the reaction for 1hr. Add 2M hydrochloric acid to the reaction solution to adjust the pH to 6~7, wash once with saturated aqueous ammonium chloride solution, extract with ethyl acetate (60mL×3), separate the liquids, combine the organic phases, wash twice with saturated brine, and anhydrous Dry over sodium sulfate and concentrate to obtain crude compound 14b. MS m/z:311.0[M+1] + .
步骤3Step 3
将化合物14b(22.62g，72.80mmol)溶于甲苯(300mL)，加入对甲苯磺酸(16.3g，94.63mmol)，100℃搅拌4hr。向反应液中加入饱和碳酸氢钠溶液调pH至中性，加入乙酸乙酯(80mL×3)萃取，分液，合并有机相，饱和食盐水洗，无水硫酸钠干燥，浓缩得产物粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100：1～3:1)得化合物14c。MS m/z:211.0[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.03(s,1H),7.69-7.63(m,1H),4.29(s,2H),4.01-3.97(m,3H)。Compound 14b (22.62g, 72.80mmol) was dissolved in toluene (300mL), p-toluenesulfonic acid (16.3g, 94.63mmol) was added, and the mixture was stirred at 100°C for 4 hours. Add saturated sodium bicarbonate solution to the reaction solution to adjust the pH to neutral, add ethyl acetate (80 mL × 3) for extraction, separate the liquids, combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~3:1) to obtain compound 14c. MS m/z:211.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.03 (s, 1H), 7.69-7.63 (m, 1H), 4.29 (s, 2H), 4.01-3.97 (m, 3H).
步骤4 Step 4
将化合物14c(1g，4.75mmol)溶于2-甲基-2-丙烯-1-醇(10mL)中，加入钠氢(284.88mg，7.12mmol，60％)，70℃反应4hr。向反应体系中加入20mL饱和氯化铵水溶液，加入乙酸乙酯(15mL×3)萃取，分液，合并有机相，饱和食盐水洗，无水硫酸钠干燥，浓缩得产物粗品。粗品经硅胶柱层析纯化(石油醚：乙酸乙酯＝100:1～1:1)得化合物14d。MS m/z:251.0[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.19(s,1H),8.68(s,1H),7.53(s,1H),5.60(s,1H),5.15(s,1H),5.12(s,1H),4.58(s,2H),1.86(s,3H)。Compound 14c (1g, 4.75mmol) was dissolved in 2-methyl-2-propen-1-ol (10mL), sodium hydrogen (284.88mg, 7.12mmol, 60%) was added, and the reaction was carried out at 70°C for 4 hours. Add 20 mL of saturated aqueous ammonium chloride solution to the reaction system, add ethyl acetate (15 mL × 3) for extraction, separate the liquids, combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1~1:1) to obtain compound 14d. MS m/z:251.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.19 (s, 1H), 8.68 (s, 1H), 7.53 (s, 1H), 5.60 (s, 1H), 5.15 (s, 1H), 5.12 (s, 1H),4.58(s,2H),1.86(s,3H).
步骤5Step 5
将化合物14d(350mg，1.40mmol)溶于二氯甲烷(5mL)，加入N,N-二异丙基乙胺(541.33mg，4.19mmol，729.56μL)，降温至0℃，加入三氟甲磺酸酐(787.85mg，2.79mmol，460.73μL)，然后在20℃下反应0.5hr。加入饱和氯化铵5mL，用乙酸乙酯(3mL×3)萃取，分液合并有机相，饱和食盐水洗，无水硫酸钠干燥，浓缩得产物粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝100:1～10:1)得化合物14e。MS m/z:382.9[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.21(s,1H),7.61(s,1H),6.96(s,1H),5.09(s,1H),5.01(s,1H),4.84(s,2H),1.85(s,3H)。Dissolve compound 14d (350 mg, 1.40 mmol) in dichloromethane (5 mL), add N,N-diisopropylethylamine (541.33 mg, 4.19 mmol, 729.56 μL), cool to 0°C, and add trifluoromethanesulfonate acid anhydride (787.85 mg, 2.79 mmol, 460.73 μL), and then reacted at 20°C for 0.5 hr. Add 5 mL of saturated ammonium chloride, extract with ethyl acetate (3 mL × 3), separate the liquids and combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100:1 ~ 10:1) to obtain compound 14e. MS m/z:382.9[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.21 (s, 1H), 7.61 (s, 1H), 6.96 (s, 1H), 5.09 (s, 1H), 5.01 (s, 1H), 4.84 (s, 2H),1.85(s,3H).
步骤6Step 6
将异丙醇(5mL)，化合物14e(430mg)加入长管中，开始搅拌；然后将化合物1c(166.88mg，盐酸盐)，N,N-二异丙基乙胺(406.55mg,3.15mmol,547.91μL)加入其中，升温至100℃，反应1小时。向反应体系中加入15mL水后，加入乙酸乙酯萃取(15mL×2)，得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝3:1～1:6)，得到化合物14f。MS m/z:382.2[M+1]+。1H NMR(400MHz,CDCl3)δppm8.84(s,1H),7.32(s,1H),6.17(s,1H),5.02(d,J＝39.6,2H),4.76-4.72(m,4H),4.37-4.34(m,2H),3.48(s,3H),3.00(s,3H),1.84(s,3H)。Add isopropyl alcohol (5mL) and compound 14e (430mg) into a long tube and start stirring; then add compound 1c (166.88mg, hydrochloride), N,N-diisopropylethylamine (406.55mg, 3.15mmol , 547.91 μL) was added, the temperature was raised to 100°C, and the reaction was carried out for 1 hour. After adding 15 mL of water to the reaction system, add ethyl acetate and extract (15 mL Get crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 3:1 to 1:6) to obtain compound 14f. MS m/z:382.2[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm8.84(s,1H),7.32(s,1H),6.17(s,1H),5.02(d,J＝39.6,2H),4.76-4.72(m,4H ),4.37-4.34(m,2H),3.48(s,3H),3.00(s,3H),1.84(s,3H).
步骤7Step 7
将二氯甲烷(6mL)，化合物14f(50mg,130.93μmol)加入拇指瓶中，开始搅拌；然后将温度降至0～5℃后，将N-碘代丁二酰亚胺(25.04mg,111.29μmol)溶于0.5mL二氯甲烷中后，缓慢滴加到其中，反应0.5小时。向反应体系中加入10mL饱和氯化铵溶液淬灭反应，再用10mL二氯甲烷萃取；有机相使用无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩。得到化合物14g，直接用于下一步。1H NMR(400MHz,CDCl3)δppm 8.71(s,1H),7.66(s,1H),5.14(s,1H),4.98(s,1H),4.84(s,2H),4.77-4.72(m,2H),4.38-4.35(m,2H),3.55-3.47(m,3H),3.01(s,3H),1.87(s,3H)。Add dichloromethane (6mL) and compound 14f (50mg, 130.93μmol) into the thumb bottle and start stirring; then lower the temperature to 0~5°C, add N-iodosuccinimide (25.04mg, 111.29 μmol) was dissolved in 0.5 mL of methylene chloride, and then slowly added dropwise to it, and the reaction was carried out for 0.5 hours. Add 10 mL of saturated ammonium chloride solution to the reaction system to quench the reaction, and then extract with 10 mL of methylene chloride; the organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C. Compound 14g was obtained and used directly in the next step. 1 H NMR (400MHz, CDCl 3 ) δppm 8.71(s,1H),7.66(s,1H),5.14(s,1H),4.98(s,1H),4.84(s,2H),4.77-4.72(m ,2H),4.38-4.35(m,2H),3.55-3.47(m,3H),3.01(s,3H),1.87(s,3H).
步骤8Step 8
将N,N-二甲基甲酰胺(5mL)，化合物14g(70mg,137.86μmol)，加入长管中，开始搅拌；氮气置换后，将甲酸钠(23.44mg,344.64μmol)，四丁基氯化铵(76.63mg,275.71μmol)，乙酸钾(54.12mg,551.43μmol)，醋酸钯(6.19mg,27.57μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入10mL水以及乙酸乙酯(10mL×2)萃取；得到的有机相用10mL饱和食盐水洗涤一次后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝1:1～1:7)，得到化合物14h。MS m/z:382.2[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.87(s,1H),7.39(s,1H),4.78-4.72(m,2H),4.39-4.33(m,4H),3.48-3.45(m,3H),2.98(s,3H),1.52(s,6H)。Add N,N-dimethylformamide (5mL) and compound 14g (70mg, 137.86μmol) into a long tube and start stirring; after nitrogen replacement, sodium formate (23.44mg, 344.64μmol), tetrabutyl chloride Ammonium (76.63 mg, 275.71 μmol), potassium acetate (54.12 mg, 551.43 μmol), and palladium acetate (6.19 mg, 27.57 μmol) were added, and the temperature was raised to 100°C and the reaction was carried out for 3 hours. Add 10 mL water and ethyl acetate (10 mL Crude. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 1:1 ~ 1:7) to obtain compound 14h. MS m/z:382.2[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.87(s,1H),7.39(s,1H),4.78-4.72(m,2H),4.39-4.33(m,4H),3.48-3.45(m,3H) ,2.98(s,3H),1.52(s,6H).
步骤9Step 9
将1,4-二氧六环(2mL)，化合物14h(13mg,34.04μmol)，化合物1h-2(6.16mg,27.23μmol)加入长管中，开始搅拌；氮气置换后，依次将乙酸钾(10.02mg,102.13μmol)，XPhos Pd G3(8.64mg,10.21μmol)加入其中，升温至100℃，反应1小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层层析法制备分离(展开剂为二氯甲烷:甲醇＝10:1)得到化合物14。 MS m/z:572.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.83(s,1H),8.11(d,J＝5.6,1H),7.81(s,1H),7.42(s,1H),6.34(d,J＝5.6,1H),4.75-4.71(m,2H),4.62-4.57(m,1H),4.49-4.44(m,1H),4.36-4.32(m,4H),3.74-3.66(m,1H),3.54-3.45(m,5H),2.98(s,3H),1.94-1.87(m,3H),1.56(d,J＝2.0,6H),1.49(d,J＝22,3H)。Add 1,4-dioxane (2mL), compound 14h (13mg, 34.04μmol), and compound 1h-2 (6.16mg, 27.23μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate ( 10.02 mg, 102.13 μmol), XPhos Pd G3 (8.64 mg, 10.21 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 1 hour. After adding 10 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure to obtain crude product. The crude product was separated by thin layer chromatography (the developing solvent was dichloromethane: methanol = 10:1) to obtain compound 14. MS m/z:572.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.83 (s, 1H), 8.11 (d, J = 5.6, 1H), 7.81 (s, 1H), 7.42 (s, 1H), 6.34 (d, J = 5.6, 1H),4.75-4.71(m,2H),4.62-4.57(m,1H),4.49-4.44(m,1H),4.36-4.32(m,4H),3.74-3.66(m,1H),3.54- 3.45(m,5H),2.98(s,3H),1.94-1.87(m,3H),1.56(d,J=2.0,6H),1.49(d,J=22,3H).
实施例15、16
Examples 15 and 16
步骤1step 1
将甲醇(100mL)，化合物15a(20g,94.76mmol)加入500mL三口瓶中，开始搅拌；再将温度升至40℃，将亚硝酸异戊酯(18.87g,161.10mmol)滴加到其中；然后将浓盐酸(4.50mL)滴加到其中，40～60℃下反应5小时。将反应液降温至0～5℃后，过滤，滤饼用3mL甲醇淋洗后，得到化合物15b。MS m/z:239.9/241.9[M+1]+。1H NMR(400MHz,DMSO-d6)δppm 12.82(s,1H),7.98(d,J＝7.6,1H),7.77(d,J＝7.6Hz,1H),7.46(d,J＝7.6Hz,1H),3.67(s,2H),3.32(s,1H)。Add methanol (100 mL) and compound 15a (20 g, 94.76 mmol) into a 500 mL three-necked flask and start stirring; then raise the temperature to 40°C, and add isoamyl nitrite (18.87 g, 161.10 mmol) dropwise into it; then Concentrated hydrochloric acid (4.50 mL) was added dropwise, and the reaction was carried out at 40 to 60°C for 5 hours. After cooling the reaction solution to 0-5°C, it was filtered, and the filter cake was rinsed with 3 mL of methanol to obtain compound 15b. MS m/z:239.9/241.9[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δppm 12.82 (s, 1H), 7.98 (d, J = 7.6, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.46 (d, J = 7.6 Hz ,1H),3.67(s,2H),3.32(s,1H).
步骤2Step 2
将1,4-二氧六环(82mL)，化合物15b(10.2g,42.49mmol)加入500mL三口瓶中，开始搅拌；然后将温度降至0～5℃，依次将三氯氧磷(19.55g,127.47mmol,11.85mL)，氯化氢1,4-二氧六环溶液(4M,5.31mL)加入其中，升温至70℃，反应6小时。反应液在45℃下进行减压浓缩，得到粗品，将该粗品与20mL乙腈混合搅拌15分钟后，过滤得滤饼即化合物15c。MS m/z:275.8/277.8[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.32(d,J＝5.6Hz,1H),8.07-8.04(m,2H),7.54(t,J＝8.0Hz,1H)。Add 1,4-dioxane (82mL) and compound 15b (10.2g, 42.49mmol) into a 500mL three-necked flask and start stirring; then lower the temperature to 0~5°C, and add phosphorus oxychloride (19.55g) , 127.47mmol, 11.85mL), hydrogen chloride 1,4-dioxane solution (4M, 5.31mL) was added, the temperature was raised to 70°C, and the reaction was carried out for 6 hours. The reaction solution was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was mixed with 20 mL of acetonitrile and stirred for 15 minutes, and then filtered to obtain a filter cake, namely compound 15c. MS m/z:275.8/277.8[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.32 (d, J = 5.6 Hz, 1H), 8.07-8.04 (m, 2H), 7.54 (t, J = 8.0 Hz, 1H).
步骤3Step 3
将1,4-二氧六环(154mL)，水(31mL)，化合物15c(7.7g,27.80mmol)，异丙烯基硼酸嚬哪醇酯(5.14g,30.58mmol)加入250mL三口瓶中，开始搅拌；氮气置换后，依次将碳酸钾(11.53g,83.41mmol)，二氯双[二叔丁基-(4-二甲基氨基苯基)膦]钯(II)(1.97g,2.78mmol,1.97mL)加入其中，升温至50℃， 反应2小时。向反应体系中加入100mL饱和氯化铵溶液以及50mL乙酸乙酯后，过滤，滤液直接分液；得到的有机相用50mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～80:20)得到化合物15d。MS m/z:237.9/239.9[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.27-8.25(m,1H),7.88(s,1H),7.66-7.59(m,2H),5.50-5.48(m,1H),5.07(s,1H),2.19(s,3H)。Add 1,4-dioxane (154mL), water (31mL), compound 15c (7.7g, 27.80mmol), isopropenyl borate zylalcohol ester (5.14g, 30.58mmol) into a 250mL three-necked flask, start Stir; after nitrogen replacement, add potassium carbonate (11.53g, 83.41mmol), dichlorobis[di-tert-butyl-(4-dimethylaminophenyl)phosphine]palladium(II) (1.97g, 2.78mmol), 1.97mL) was added, and the temperature was raised to 50°C. Reaction takes 2 hours. Add 100 mL of saturated ammonium chloride solution and 50 mL of ethyl acetate to the reaction system, filter, and separate the filtrate directly; wash the obtained organic phase with 50 mL of saturated brine, add anhydrous sodium sulfate, dry, filter, and keep the filtrate at 45°C Concentrate under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~80:20) to obtain compound 15d. MS m/z:237.9/239.9[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.27-8.25(m,1H),7.88(s,1H),7.66-7.59(m,2H),5.50-5.48(m,1H),5.07(s,1H) ,2.19(s,3H).
步骤4Step 4
将四氢呋喃(130mL)，化合物15d(5.40g,22.7mmol)加入250mL三口瓶中，开始搅拌；氮气置换后将四甲基乙二胺(3.95g,34.0mmol,5.13mL)，1,1-双(二苯基磷)二茂铁氯化钯(332mg,454μmol)加入其中；然后将硼氢化钠(1.84g,48.8mmol)缓慢加到其中，室温20℃，反应4小时。将反应液缓慢倒入50mL盐酸(1M)淬灭，然后加入乙酸乙酯萃取(25mL×2)，得到的有机相用50mL饱和食盐水洗涤一次后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～80:20)得到化合物15e。MS m/z:204.0[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),7.93(s,1H),7.90-7.88(m,1H),7.59-7.56(m,2H),5.49(m,1H),5.08(s,1H),2.21(s,3H)。Add tetrahydrofuran (130mL) and compound 15d (5.40g, 22.7mmol) into a 250mL three-necked flask and start stirring; after nitrogen replacement, add tetramethylethylenediamine (3.95g, 34.0mmol, 5.13mL), 1,1-bis (Diphenylphosphorus)ferrocene palladium chloride (332 mg, 454 μmol) was added; then sodium borohydride (1.84 g, 48.8 mmol) was slowly added to the mixture, and the reaction was carried out at room temperature 20°C for 4 hours. Slowly pour the reaction solution into 50 mL hydrochloric acid (1M) to quench, then add ethyl acetate for extraction (25 mL Concentrate under reduced pressure at 45°C. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~80:20) to obtain compound 15e. MS m/z:204.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07(s,1H),7.93(s,1H),7.90-7.88(m,1H),7.59-7.56(m,2H),5.49(m,1H),5.08 (s,1H),2.21(s,3H).
步骤5Step 5
将乙酸乙酯(50mL)，二氧化铂(189.54mg,834.70μmol)加入5mL拇指瓶中，开始搅拌；然后将化合物15e(3.4g,16.69mmol)加入其中，氢气置换后，压力15Psi，室温20℃反应15小时。反应液直接过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～90:10)得到化合物15f。MS m/z:206.1[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.06(s,1H),7.94(s,1H),7.82(d,J＝8.0Hz,1H),7.65-7.64(m,1H),7.59-7.55(m,1H),3.66-3.55(m,1H),1.40(d,J＝6.8Hz,6H)。Add ethyl acetate (50mL) and platinum dioxide (189.54mg, 834.70μmol) into a 5mL thumb bottle and start stirring; then add compound 15e (3.4g, 16.69mmol) into it. After hydrogen replacement, the pressure is 15Psi and room temperature is 20 ℃ reaction for 15 hours. The reaction solution was directly filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~90:10) to obtain compound 15f. MS m/z:206.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.06 (s, 1H), 7.94 (s, 1H), 7.82 (d, J = 8.0Hz, 1H), 7.65-7.64 (m, 1H), 7.59-7.55 (m ,1H),3.66-3.55(m,1H),1.40(d,J=6.8Hz,6H).
步骤6Step 6
将硫酸(66mL,95％纯度)，化合物15f(3.3g,16.04mmol)加入250mL三口瓶中，开始搅拌；然后将N-溴代丁二酰亚胺(4.28g,24.07mmol)加入其中，升温至60℃反应5小时。将反应液缓慢加入300mL冰水中后，加入固体氢氧化钠将pH值调至8～9；然后加入乙酸乙酯萃取(100mL×2)，得到的有机相加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～90:10)得到化合物15g。MS m/z:283.8/285.8[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.45(s,1H),7.93(s,1H),7.79(d,J＝8.0Hz,1H),7.47(d,J＝7.6Hz,1H),3.63-3.52(m,1H),1.39(d,J＝6.8Hz,6H)。Add sulfuric acid (66 mL, 95% purity) and compound 15f (3.3 g, 16.04 mmol) into a 250 mL three-neck flask and start stirring; then add N-bromosuccinimide (4.28 g, 24.07 mmol) and raise the temperature. React at 60°C for 5 hours. After slowly adding the reaction solution to 300 mL of ice water, add solid sodium hydroxide to adjust the pH value to 8-9; then add ethyl acetate for extraction (100 mL × 2). The obtained organic phase is dried with anhydrous sodium sulfate, filtered, and the filtrate Concentrate under reduced pressure at 45°C. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~90:10) to obtain 15g of compound. MS m/z:283.8/285.8[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.45 (s, 1H), 7.93 (s, 1H), 7.79 (d, J = 8.0Hz, 1H), 7.47 (d, J = 7.6Hz, 1H), 3.63 -3.52(m,1H),1.39(d,J＝6.8Hz,6H).
步骤7Step 7
将1,4二氧六环(6mL)，化合物15g(0.2g,702.79μmol)，化合物9b的盐酸盐(178.55mg)加入长管中，开始搅拌；氮气置换后，依次将碳酸铯(915.94mg,2.81mmol)，Xantphos Pd G4(67.64mg,70.28μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入10mL饱和氯化铵溶液以及10mL乙酸乙酯后，分液；得到的水相用乙酸乙酯10mL萃取后，合并有机相，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝5:1～1:3)得到化合物15h。MS m/z:379.0[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.12(s,1H),7.85(s,1H),7.48(d,J＝8.0Hz,1H),6.53(d,J＝8.0Hz,1H),4.45-4.41(m,2H),4.16-3.98(m,2H),3.50-3.42(m,2H),3.24-3.01(m,3H),2.51-2.43(m,1H),2.33-2.11(m,2H),1.96-1.86(m,1H),1.34(d,J＝7.2Hz,6H)。Add 1,4 dioxane (6mL), compound 15g (0.2g, 702.79μmol), and the hydrochloride of compound 9b (178.55mg) into a long tube, and start stirring; after nitrogen replacement, add cesium carbonate (915.94 mg, 2.81 mmol), Xantphos Pd G4 (67.64 mg, 70.28 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. After adding 10 mL of saturated ammonium chloride solution and 10 mL of ethyl acetate to the reaction system, separate the liquids; extract the obtained aqueous phase with 10 mL of ethyl acetate, combine the organic phases, add anhydrous sodium sulfate, dry, filter, and the filtrate is at 45 The crude product was obtained by concentrating under reduced pressure at ℃. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1~1:3) to obtain compound 15h. MS m/z:379.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.12 (s, 1H), 7.85 (s, 1H), 7.48 (d, J = 8.0Hz, 1H), 6.53 (d, J = 8.0Hz, 1H), 4.45- 4.41(m,2H),4.16-3.98(m,2H),3.50-3.42(m,2H),3.24-3.01(m,3H),2.51-2.43(m,1H),2.33-2.11(m,2H ), 1.96-1.86 (m, 1H), 1.34 (d, J = 7.2Hz, 6H).
步骤8Step 8
将1,4二氧六环(4mL)，化合物15h(80mg,211.13μmol)，化合物1h-2(43.95mg,194.24μmol)加入长管中，开始搅拌；氮气置换后，将碳酸铯(171.98mg,527.82μmol)，BrettPhos Pd G3(38.28mg,42.23μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；有机相用饱和食盐水(10mL)洗涤一次后，无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝1:1～0:1)，得到产品SFC拆分(色谱柱：Phenomenex- Cellulose-2(250mm×30mm，10μm)；流动相：A(超临界CO2)和B(乙醇，含0.1％氨水)；梯度：B％＝55％-55％)分离得到化合物15和化合物16。Add 1,4 dioxane (4mL), compound 15h (80mg, 211.13μmol), and compound 1h-2 (43.95mg, 194.24μmol) into a long tube and start stirring; after nitrogen replacement, add cesium carbonate (171.98mg , 527.82 μmol), BrettPhos Pd G3 (38.28 mg, 42.23 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. After adding 10 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1 ~ 0:1), and the product was obtained for SFC separation (chromatography column: Phenomenex- Cellulose-2 (250mm×30mm, 10μm); mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% ammonia); gradient: B% = 55%-55%) to separate compound 15 and compound 16 .
化合物15：MS m/z:569.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.51(s,1H),8.08(d,J＝5.6Hz,1H),7.43-7.37(m,2H),6.41(d,J＝7.6Hz,1H),6.23(d,J＝5.6Hz,1H),4.63-4.58(m,2H),4.49-4.38(m,3H),4.18-4.14(m,1H),3.99-3.95(m,1H),3.75-3.66(m,1H),3.61-3.44(m,4H),3.24-3.01(m,3H),2.52-2.44(m,1H),2.32-2.10(m,2H),2.05-1.88(m,3H),1.51(d,J＝21.6,3H),1.36(d,J＝6.8Hz,6H)。SFC(色谱柱：Lux Cellulose-2，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(甲醇，含0.1％异丙胺)；梯度：B％＝50～50％，5min；流速:4.0mL/min；波长:220nm；压力:1800psi)，Rt＝2.389min，ee：100％。Compound 15: MS m/z: 569.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07 (s, 1H), 8.51 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.43-7.37 (m, 2H), 6.41 (d, J ＝7.6Hz,1H),6.23(d,J＝5.6Hz,1H),4.63-4.58(m,2H),4.49-4.38(m,3H),4.18-4.14(m,1H),3.99-3.95( m,1H),3.75-3.66(m,1H),3.61-3.44(m,4H),3.24-3.01(m,3H),2.52-2.44(m,1H),2.32-2.10(m,2H), 2.05-1.88(m,3H),1.51(d,J＝21.6,3H),1.36(d,J＝6.8Hz,6H). SFC (column: Lux Cellulose-2, 3μm, 0.46cm id×5cm L; mobile phase: A (supercritical CO 2 ) and B (methanol, containing 0.1% isopropylamine); gradient: B%=50~50% , 5min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi), Rt=2.389min, ee: 100%.
化合物16：MS m/z:569.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.50(s,1H),8.08(d,J＝5.6Hz,1H),7.43-7.41(m,2H),6.42(d,J＝8.0Hz,1H),6.30-6.19(m,1H),4.67-4.37(m,4H),4.18-3.96(m,2H),3.78-3.41(m,5H),3.28-2.98(m,3H),2.51-2.13(m,3H),1.98-1.89(m,4H),1.52(d,J＝21.6,3H),1.36(d,J＝6.8Hz,6H)。SFC(色谱柱：Lux Cellulose-2，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(甲醇，含0.1％异丙胺)；梯度：B％＝50～50％，5min；流速:4.0mL/min；波长:220nm；压力:1800psi)，Rt＝3.136min,ee：99.06％。Compound 16: MS m/z: 569.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07 (s, 1H), 8.50 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.43-7.41 (m, 2H), 6.42 (d, J ＝8.0Hz,1H),6.30-6.19(m,1H),4.67-4.37(m,4H),4.18-3.96(m,2H),3.78-3.41(m,5H),3.28-2.98(m,3H) ),2.51-2.13(m,3H),1.98-1.89(m,4H),1.52(d,J＝21.6,3H),1.36(d,J＝6.8Hz,6H). SFC (column: Lux Cellulose-2, 3μm, 0.46cm id×5cm L; mobile phase: A (supercritical CO 2 ) and B (methanol, containing 0.1% isopropylamine); gradient: B%=50~50% , 5min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi), Rt=3.136min, ee: 99.06%.
实施例17、18
Examples 17 and 18
步骤1step 1
将化合物17a(590mg,2.33mmol)溶于DCM(15mL)，0℃下缓慢加入Dess-Martin试剂(1.48g,3.49mmol,91.72μL)，后升至25℃搅拌反应2小时。反应液过滤，滤液减压浓缩得到化合物17b。1H NMR(400MHz,CDCl3)δppm 7.58-7.48(m,4H),7.38-7.12(m,6H),4.62(s,1H),4.20(dd,J＝19.01,10.38Hz,1H),4.07(dd,J＝14.45,7.07Hz,1H),3.75(d,J＝4.8Hz,1H),0.97-0.92(d,J＝4.0Hz,3H)。Compound 17a (590 mg, 2.33 mmol) was dissolved in DCM (15 mL), Dess-Martin reagent (1.48 g, 3.49 mmol, 91.72 μL) was slowly added at 0°C, and then raised to 25°C and stirred for 2 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 17b. 1 H NMR (400MHz, CDCl 3 ) δppm 7.58-7.48 (m, 4H), 7.38-7.12 (m, 6H), 4.62 (s, 1H), 4.20 (dd, J = 19.01, 10.38Hz, 1H), 4.07 (dd, J=14.45, 7.07Hz, 1H), 3.75 (d, J=4.8Hz, 1H), 0.97-0.92 (d, J=4.0Hz, 3H).
步骤2Step 2
将环丁砜(191.26mg,1.59mmol)溶解于THF(5mL)，氮气保护下，温度降至-70℃，将正丁基锂(2.5M,636.63μL)滴加至反应液中，搅拌反应1小时，将化合物17b(200mg,795.79μmol)加入至反应液中， 继续反应1小时。用3mL饱和氯化铵溶液淬灭反应，用乙酸乙酯(20mL)萃取，合并有机相并用无水硫酸钠干燥后减压浓缩得到粗品，粗品经制备高效液相色谱(色谱柱:Welch Xtimate C18 100×40mm×3μm；流动相:[水(三氟乙酸)-乙腈]；乙腈％:10％-40％,8min)纯化得到化合物17c。MS m/z:372.1[M+1]+。Dissolve sulfolane (191.26mg, 1.59mmol) in THF (5mL). Under nitrogen protection, lower the temperature to -70°C. Add n-butyllithium (2.5M, 636.63μL) dropwise into the reaction solution and stir for 1 hour. , add compound 17b (200mg, 795.79μmol) into the reaction solution, Continue the reaction for 1 hour. The reaction was quenched with 3 mL of saturated ammonium chloride solution, extracted with ethyl acetate (20 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product, which was subjected to preparative high-performance liquid chromatography (column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile%: 10%-40%, 8min) to obtain compound 17c. MS m/z:372.1[M+1] + .
步骤3Step 3
在反应瓶中加入化合物17c(400mg,1.08mmol)，二氯甲烷(10mL)，氮气置换三次降温至-20℃加入二乙氨基三氟化硫(347.12mg,2.15mmol,284μL)自然升温至25℃搅拌3小时。反应液缓慢加入到碳酸氢钠水溶液(10mL)中淬灭，然后用二氯甲烷萃取(20mL×2)，收集有机相，有机相用饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品用柱层析纯化(石油醚：乙酸乙酯＝10:0-1:1)纯化得到化合物17d。MS m/z:374.3[M+1]+。1H NMR(400MHz,CDCl3)δppm 7.42(m,6H),7.16-7.33(m,4H),4.49(s,1H),3.80(dd,J＝19.01,10.38Hz,1H),3.46(dd,J＝14.45,7.07Hz,1H),3.23-3.40(m,2H),3.04-3.12(m,1H),2.88-3.01(m,1H),2.30-2.40(m,1H),2.21-2.29(m,1H),2.09-2.18(m,2H),0.86(dd,J＝6.44,1.19Hz,3H)。Add compound 17c (400 mg, 1.08 mmol) and dichloromethane (10 mL) to the reaction flask, replace with nitrogen three times and cool the temperature to -20°C. Add diethylaminosulfur trifluoride (347.12 mg, 2.15 mmol, 284 μL) and naturally raise the temperature to 25 °C and stirred for 3 hours. The reaction solution was slowly added to aqueous sodium bicarbonate solution (10 mL) to quench, and then extracted with dichloromethane (20 mL × 2). The organic phase was collected, washed with saturated brine (10 mL × 2), and dried over anhydrous sodium sulfate. , filtered, and concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:0-1:1) to obtain compound 17d. MS m/z:374.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 7.42 (m, 6H), 7.16-7.33 (m, 4H), 4.49 (s, 1H), 3.80 (dd, J = 19.01, 10.38Hz, 1H), 3.46 (dd ,J＝14.45,7.07Hz,1H),3.23-3.40(m,2H),3.04-3.12(m,1H),2.88-3.01(m,1H),2.30-2.40(m,1H),2.21-2.29 (m,1H),2.09-2.18(m,2H),0.86(dd,J=6.44,1.19Hz,3H).
步骤4Step 4
在反应瓶中加入化合物17d(100mg,267.75μmol)，甲醇(2mL)，盐酸(12M,44.62μL)，氢氧化钯(30.00mg,213.61μmol)氢气置换三次，25℃，15psi搅拌16小时反应液进行过滤，滤液减压浓缩得到化合物17j。1H NMR(400MHz,CD3OD)δppm 4.78-4.83(m,1H),4.43-4.60(m,1H),4.18-4.33(m,1H),3.69-3.84(m,1H),3.14-3.25(m,1H),2.94-3.08(m,1H),2.41-2.54(m,1H),2.21-2.35(m,1H),2.06-2.21(m,2H),1.55-1.62(m,3H)。Add compound 17d (100 mg, 267.75 μmol), methanol (2 mL), hydrochloric acid (12 M, 44.62 μL), and palladium hydroxide (30.00 mg, 213.61 μmol) into the reaction bottle. Replace it with hydrogen three times. Stir the reaction solution at 25°C and 15 psi for 16 hours. Filtration was performed, and the filtrate was concentrated under reduced pressure to obtain compound 17j. 1 H NMR (400MHz, CD 3 OD) δppm 4.78-4.83(m,1H),4.43-4.60(m,1H),4.18-4.33(m,1H),3.69-3.84(m,1H),3.14-3.25 (m,1H),2.94-3.08(m,1H),2.41-2.54(m,1H),2.21-2.35(m,1H),2.06-2.21(m,2H),1.55-1.62(m,3H) .
步骤5Step 5
在反应瓶中加入化合物17j(80mg,41.03μmol)，化合物15g(93.38mg,41.03μmol)，二氧六环(2mL)，碳酸铯(427.78mg,164.12μmol)氮气置换三次加入Xantphos Pd G4(63.18mg,8.21μmol)，100℃搅拌2hr。反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品，再通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物17k。MS m/z:411.1[M+1]+。Add compound 17j (80 mg, 41.03 μmol), compound 15g (93.38 mg, 41.03 μmol), dioxane (2 mL), and cesium carbonate (427.78 mg, 164.12 μmol) to the reaction bottle. Nitrogen replacement was performed three times and Xantphos Pd G4 (63.18 mg, 8.21 μmol), stir at 100°C for 2 hr. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was then passed through the column layer. Compound 17k was purified by analytical purification (dichloromethane: methanol = 10:0-10:1). MS m/z:411.1[M+1] + .
步骤6Step 6
在反应瓶中加入化合物17k(100mg,243.35μmol)，化合物17i-2(55.06mg,243.35μmol)，二氧六环(2mL)，氮气置换三次加入BrettPhos Pd G3(44.12mg,48.67μmol)，乙酸钾(71.65mg,730.05μmol)，100℃搅拌3小时。反应完成后，反应液进行过滤，滤液进行减压浓缩得到粗品。粗品经制备高效液相色谱分离(色谱柱:Waters Xbridge BEH C18 100×30mm×10μm；流动相:A(乙腈)和B(水，含0.05％碳酸氢铵)；梯度：B％:35％-60％，8min)后，送SFC拆分(色谱柱：ChiralPak IH,250×30mm,10μm；流动相：A(超临界CO2)和B(甲醇，含0.1％氨水)；梯度：B％＝70％-70％，40min)得到化合物17和18。Add compound 17k (100 mg, 243.35 μmol), compound 17i-2 (55.06 mg, 243.35 μmol), dioxane (2 mL) to the reaction bottle, replace with nitrogen three times and add BrettPhos Pd G3 (44.12 mg, 48.67 μmol), acetic acid Potassium (71.65 mg, 730.05 μmol), stir at 100°C for 3 hours. After the reaction is completed, the reaction solution is filtered, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Waters Xbridge BEH C18 100×30mm×10μm; mobile phase: A (acetonitrile) and B (water, containing 0.05% ammonium bicarbonate); gradient: B%: 35%- 60%, 8min), send to SFC for separation (chromatographic column: ChiralPak IH, 250×30mm, 10μm; mobile phase: A (supercritical CO 2 ) and B (methanol, containing 0.1% ammonia); gradient: B%＝ 70%-70%, 40min) to obtain compounds 17 and 18.
化合物17：MS m/z:601.2[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.02(s,1H),8.54(s,1H),8.08(d,J＝5.70Hz,1H),7.54(dd,J＝4.28,1.64Hz,1H),7.42(d,J＝7.89Hz,1H),6.52(d,J＝8.11Hz,1H),6.25(dt,J＝2.69,1.40Hz,1H),4.64(d,J＝9.65Hz,4H),4.44-4.53(m,1H),4.05-4.32(m,1H),3.67-3.79(m,1H),3.52-3.61(m,3H),3.13-3.23(m,1H),3.07(dd,J＝12.17,6.91Hz,1H),2.14-2.41(m,2H),1.81-2.01(m,2H),1.49-1.55(m,6H),1.37(dd,J＝9.87,6.80Hz,9H)。SFC(色谱柱：Chiralpak IH-3,50×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(甲醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:254nm；压力:1800psi)，Rt＝1.309min，ee：100％。Compound 17: MS m/z: 601.2[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.02 (s, 1H), 8.54 (s, 1H), 8.08 (d, J = 5.70Hz, 1H), 7.54 (dd, J = 4.28, 1.64Hz, 1H), 7.42(d,J＝7.89Hz,1H),6.52(d,J＝8.11Hz,1H),6.25(dt,J＝2.69,1.40Hz,1H),4.64(d,J＝9.65Hz,4H), 4.44-4.53(m,1H),4.05-4.32(m,1H),3.67-3.79(m,1H),3.52-3.61(m,3H),3.13-3.23(m,1H),3.07(dd,J ＝12.17,6.91Hz,1H),2.14-2.41(m,2H),1.81-2.01(m,2H),1.49-1.55(m,6H),1.37(dd,J＝9.87,6.80Hz,9H). SFC (column: Chiralpak IH-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (methanol, containing 0.1% isopropylamine); gradient: B%=50~50%, 4min; flow rate: 4.0mL/min; wavelength: 254nm; pressure: 1800psi), Rt=1.309min, ee: 100%.
化合物18：MS m/z:601.2[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.03(s,1H),8.37-8.65(m,1H),8.09(d,J＝5.25Hz,1H),7.43(d,J＝7.88Hz,1H),6.53(d,J＝7.75Hz,1H),6.20-6.35(m,1H),4.43-4.74(m,4H),4.21-4.31(m,1H),3.68-3.83(m,1H),3.51-3.64(m,4H),3.13-3.23(m,1H),2.95-3.10(m,1H),2.46-2.57(m,1H),2.15-2.40(m,4H),1.89-2.07(m,3H),1.49-1.55(m,3H),1.32-1.40(m,9H)。SFC(色谱柱：Chiralpak IH-3,50×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(甲醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:254nm；压力:1800psi)，Rt＝2.628min，ee：100％。 Compound 18: MS m/z: 601.2[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.03 (s, 1H), 8.37-8.65 (m, 1H), 8.09 (d, J = 5.25Hz, 1H), 7.43 (d, J = 7.88Hz, 1H), 6.53(d,J＝7.75Hz,1H),6.20-6.35(m,1H),4.43-4.74(m,4H),4.21-4.31(m,1H),3.68-3.83(m,1H),3.51- 3.64(m,4H),3.13-3.23(m,1H),2.95-3.10(m,1H),2.46-2.57(m,1H),2.15-2.40(m,4H),1.89-2.07(m,3H ),1.49-1.55(m,3H),1.32-1.40(m,9H). SFC (column: Chiralpak IH-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (methanol, containing 0.1% isopropylamine); gradient: B%=50~50%, 4min; flow rate: 4.0mL/min; wavelength: 254nm; pressure: 1800psi), Rt=2.628min, ee: 100%.
实施例19、20
Examples 19 and 20
步骤1step 1
在100mL三口瓶中加入化合物17a(3g,11.84mmol)，二氯甲烷(50mL)，三乙胺(1.44g,14.21mmol,1.98mL)，氮气置换三次，降温至0℃滴入甲烷磺酰氯(1.49g,13.03mmol,1.01mL)，自然升温至25℃搅拌4小时。反应液缓慢加入到冰水溶液(50mL)中淬灭，然后用二氯甲烷萃取(50mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝1:0～1:1)纯化得到化合物19g。1H NMR(400MHz,CDCl3)δppm 7.32-7.42(m,4H),7.13-7.30(m,6H),4.59(d,J＝6.13Hz,1H),4.38(s,1H),3.71(t,J＝7.32Hz,1H),3.36(t,J＝6.13Hz,1H),2.95(s,3H),2.85(dd,J＝7.94,6.82Hz,1H),0.79(d,J＝6.25Hz,3H)。Add compound 17a (3g, 11.84mmol), dichloromethane (50mL), and triethylamine (1.44g, 14.21mmol, 1.98mL) into a 100mL three-necked flask, replace with nitrogen three times, cool to 0°C and drop in methanesulfonyl chloride ( 1.49g, 13.03mmol, 1.01mL), naturally raise the temperature to 25°C and stir for 4 hours. The reaction solution was slowly added to ice-water solution (50 mL) to quench, and then extracted with dichloromethane (50 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 1:0-1:1) to obtain compound 19g. 1 H NMR (400MHz, CDCl 3 ) δppm 7.32-7.42 (m, 4H), 7.13-7.30 (m, 6H), 4.59 (d, J = 6.13Hz, 1H), 4.38 (s, 1H), 3.71 (t ,J＝7.32Hz,1H),3.36(t,J＝6.13Hz,1H),2.95(s,3H),2.85(dd,J＝7.94,6.82Hz,1H),0.79(d,J＝6.25Hz ,3H).
步骤2Step 2
在100mL三口反应瓶中加入化合物19a(10g,83.22mmol,7.94mL)，四氢呋喃(300mL)，氮气置换三次，降温至-65℃滴入二(三甲基硅基)氨基锂(1M,166.43mL)，-65℃搅拌0.5hr，加入氯甲酸甲酯(7.86g,83.22mmol,6.45mL)，-65℃继续反应2.5小时。反应液缓慢加入到氯化铵水溶液(100mL)中淬灭，然后用乙酸乙酯萃取(200mL×2)，收集有机相，有机相用饱和食盐水(200mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品，粗品通过柱层析纯化(石油醚：乙酸乙酯＝1:0-1:1)纯化得到化合物19b。1H NMR(400MHz,CDCl3)δppm 3.93(t,J＝7.69Hz,1H),3.85(s,3H),3.10-3.17(m,2H),3.01-3.05(m,1H),2.52-2.61(m,1H),2.34-2.42(m,1H),2.20-2.26(m,1H)。 Add compound 19a (10g, 83.22mmol, 7.94mL) and tetrahydrofuran (300mL) to a 100mL three-neck reaction flask, replace with nitrogen three times, cool to -65°C and drop in lithium bis(trimethylsilyl)amide (1M, 166.43mL). ), stir at -65°C for 0.5hr, add methyl chloroformate (7.86g, 83.22mmol, 6.45mL), and continue the reaction at -65°C for 2.5 hours. The reaction solution was slowly added to aqueous ammonium chloride solution (100 mL) to quench, and then extracted with ethyl acetate (200 mL × 2). The organic phase was collected, washed with saturated brine (200 mL × 2), and dried over anhydrous sodium sulfate. , filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 1:0-1:1) to obtain compound 19b. 1 H NMR (400MHz, CDCl 3 ) δppm 3.93 (t, J = 7.69Hz, 1H), 3.85 (s, 3H), 3.10-3.17 (m, 2H), 3.01-3.05 (m, 1H), 2.52-2.61 (m,1H),2.34-2.42(m,1H),2.20-2.26(m,1H).
步骤3Step 3
在反应瓶中加入化合物19b(0.6g,1.81mmol)，化合物19g(419.40mg,2.35mmol)，N,N-二甲基甲酰胺(10mL)，氮气置换三次，加入氢化钠(86.90mg,2.17mmol,60％纯度)，25℃搅拌15分钟升温至80℃搅拌1小时。反应液缓慢加入到氯化铵水溶液(20mL)中淬灭后，使用乙酸乙酯萃取(30mL×2)，无水硫酸钠干燥，过滤，减压浓缩得到粗品，粗品通过柱层析纯化(石油醚：乙酸乙酯＝10:0-1:1)纯化得到化合物19c。1H NMR(400MHz,CDCl3)δppm 7.39(dd,J＝19.39,7.13Hz,5H),7.28-7.32(m,2H),7.24(s,3H),4.42(s,1H),3.80(s,2H),3.55-3.67(m,1H),3.09-3.22(m,3H),2.80-2.94(m,2H),2.65-2.76(m,1H),2.34-2.48(m,1H),2.20-2.32(m,1H),2.07-2.16(m,1H),1.25-1.39(m,1H),0.73(d,J＝5.63Hz,3H)。Add compound 19b (0.6g, 1.81mmol), compound 19g (419.40mg, 2.35mmol), N,N-dimethylformamide (10mL) into the reaction bottle, replace with nitrogen three times, and add sodium hydride (86.90mg, 2.17 mmol, 60% purity), stir at 25°C for 15 minutes, increase the temperature to 80°C and stir for 1 hour. The reaction solution was slowly added to an aqueous ammonium chloride solution (20 mL) to quench, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum Ether: ethyl acetate = 10:0-1:1) was purified to obtain compound 19c. 1 H NMR (400MHz, CDCl 3 ) δppm 7.39 (dd, J＝19.39, 7.13Hz, 5H), 7.28-7.32 (m, 2H), 7.24 (s, 3H), 4.42 (s, 1H), 3.80 (s ,2H),3.55-3.67(m,1H),3.09-3.22(m,3H),2.80-2.94(m,2H),2.65-2.76(m,1H),2.34-2.48(m,1H),2.20 -2.32(m,1H),2.07-2.16(m,1H),1.25-1.39(m,1H),0.73(d,J=5.63Hz,3H).
步骤4Step 4
在反应瓶中加入化合物19c(250mg,604.55μmol)，N,N-二甲基乙酰胺(5mL)，氯化锂(205.02mg,4.84mmol,99.04μL)，氮气置换三次，140℃搅拌10小时。反应液冷却至室温后，反应液缓慢加入到水溶液(10mL)中，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用饱和食盐水洗涤(10mL×2)，无水硫酸钠干燥，过滤，减压浓缩得到粗品，粗品通过柱层析纯化(石油醚：乙酸乙酯＝1:0-1:1)得到化合物19d。MS m/z:356.3[M+1]+。Add compound 19c (250 mg, 604.55 μmol), N,N-dimethylacetamide (5 mL), and lithium chloride (205.02 mg, 4.84 mmol, 99.04 μL) into the reaction bottle, replace with nitrogen three times, and stir at 140°C for 10 hours. . After the reaction solution was cooled to room temperature, the reaction solution was slowly added to the aqueous solution (10 mL), and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, and the organic phase was washed with saturated brine (10 mL × 2), anhydrous sulfuric acid Dried over sodium, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 1:0-1:1) to obtain compound 19d. MS m/z:356.3[M+1] + .
步骤5Step 5
在反应瓶中加入化合物19d(100mg,281.30μmol)，甲醇(1mL)，盐酸(12M,46.88μL,2eq)，氢氧化钯(30.00mg,42.72μmol,20％纯度)，氢气置换三次，50℃，15psi，搅拌10小时。反应液进行过滤，滤液减压浓缩得到化合物19e的盐酸盐。1H NMR(400MHz,CD3OD)δppm 4.41-4.55(m,1H),3.96-4.10(m,2H),3.38-3.46(m,1H),3.12-3.25(m,1H),2.87-3.09(m,2H),2.30-2.49(m,1H),2.18-2.28(m,1H),2.03-2.16(m,1H),1.80(m,1H),1.59(dd,J＝17.20,6.69Hz,3H)。Add compound 19d (100mg, 281.30μmol), methanol (1mL), hydrochloric acid (12M, 46.88μL, 2eq), palladium hydroxide (30.00mg, 42.72μmol, 20% purity) into the reaction bottle, replace with hydrogen three times, 50°C , 15psi, stir for 10 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the hydrochloride of compound 19e. 1 H NMR (400MHz, CD 3 OD) δppm 4.41-4.55(m,1H),3.96-4.10(m,2H),3.38-3.46(m,1H),3.12-3.25(m,1H),2.87-3.09 (m,2H),2.30-2.49(m,1H),2.18-2.28(m,1H),2.03-2.16(m,1H),1.80(m,1H),1.59(dd,J＝17.20,6.69Hz ,3H).
步骤6Step 6
在反应瓶中加入化合物19e的盐酸盐(50mg)，化合物15g(63.03mg,221.50μmol)，二氧六环(2mL)，碳酸铯(216.50mg,664.49μmol)，氮气置换三次加入Xantphos-Pd-G4(31.97mg,33.22μmol)，100℃搅拌8小时。反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)得到化合物19f。Add the hydrochloride of compound 19e (50 mg), compound 15 g (63.03 mg, 221.50 μmol), dioxane (2 mL), and cesium carbonate (216.50 mg, 664.49 μmol) into the reaction flask, replace with nitrogen three times and add Xantphos-Pd -G4 (31.97 mg, 33.22 μmol), stir at 100°C for 8 hours. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 19f.
步骤7Step 7
在反应瓶中加入化合物19f(120mg,305.39μmol)，化合物1h-2(76.00mg,335.93μmol)，二氧六环(2mL)，氮气置换三次加入BrettPhos Pd G3(55.37mg,61.08μmol)，碳酸铯(298.51mg,916.17μmol)，100℃搅拌3小时。反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化后，经SFC拆分(色谱柱：DAICEL CHIRALPAK IC(250mm×30mm,10μm)；流动相：A(超临界CO2)和B(乙醇，含0.1％氨水)；梯度：B％＝64％-64％，20min)得到化合物19和化合物20。Add compound 19f (120 mg, 305.39 μmol), compound 1h-2 (76.00 mg, 335.93 μmol), dioxane (2 mL) to the reaction bottle, replace with nitrogen three times and add BrettPhos Pd G3 (55.37 mg, 61.08 μmol), carbonic acid Cesium (298.51 mg, 916.17 μmol), stirred at 100°C for 3 hours. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane: methanol = 10:0-10:1) and then separated by SFC (chromatographic column: DAICEL CHIRALPAK IC (250mm×30mm, 10μm); mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% ammonia); gradient: B% = 64%-64%, 20 min) to obtain compound 19 and compound 20.
化合物19：MS m/z:583.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.08(s,1H),8.37-8.64(m,1H),8.12-8.26(m,1H),8.09(d,J＝6.25Hz,1H),7.47(d,J＝8.00Hz,1H),6.58(d,J＝8.00Hz,1H),6.23-6.48(m,1H),4.66-4.77(m,1H),4.61(m,2H),4.44(m,1H),3.69-3.79(m,1H),3.47-3.67(m,4H),3.28-3.37(m,1H),3.16-3.25(m,1H),3.00-3.09(m,1H),2.81-2.91(m,1H),2.22-2.38(m,2H),2.09-2.19(m,1H),1.93-2.07(m,3H),1.80-1.88(m,1H),1.50-1.58(m,6H),1.36(t,J＝7.25Hz,6H)。SFC检测(色谱柱：Chiralpak IC-3,50×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi)，Rt＝0.941min，ee：100％。Compound 19: MS m/z: 583.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.08 (s, 1H), 8.37-8.64 (m, 1H), 8.12-8.26 (m, 1H), 8.09 (d, J = 6.25Hz, 1H), 7.47 (d ,J＝8.00Hz,1H),6.58(d,J＝8.00Hz,1H),6.23-6.48(m,1H),4.66-4.77(m,1H),4.61(m,2H),4.44(m, 1H),3.69-3.79(m,1H),3.47-3.67(m,4H),3.28-3.37(m,1H),3.16-3.25(m,1H),3.00-3.09(m,1H),2.81- 2.91(m,1H),2.22-2.38(m,2H),2.09-2.19(m,1H),1.93-2.07(m,3H),1.80-1.88(m,1H),1.50-1.58(m,6H ), 1.36 (t, J = 7.25Hz, 6H). SFC detection (chromatographic column: Chiralpak IC-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~50% , 4min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi), Rt=0.941min, ee: 100%.
化合物20：MS m/z:583.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.09(s,1H),8.43-8.55(m,1H),8.08(d,J＝5.88Hz,1H),7.76-7.97(m,1H),7.43(d,J＝7.88Hz,1H),6.52(d,J＝8.00Hz,1H),6.21-6.41(m,1H),4.61-4.75 (m,2H),4.45-4.57(m,1H),4.24(t,J＝6.00Hz,1H),3.82(t,J＝7.07Hz,1H),3.68-3.78(m,1H),3.49-3.64(m,3H),3.25-3.34(m,1H),3.14-3.22(m,1H),3.04(dt,J＝13.20,8.79Hz,1H),2.72-2.82(m,1H),2.41-2.54(m,1H),2.24-2.35(m,1H),2.11-2.21(m,1H),2.06(m,1H),1.88-2.01(m,3H),1.48-1.58(m,3H),1.46(d,J＝6.13Hz,3H),1.36(t,J＝7.44Hz,6H)。SFC(色谱柱：Chiralpak IC-3,50×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi)，Rt＝1.411min，ee：100％。Compound 20: MS m/z: 583.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.09 (s, 1H), 8.43-8.55 (m, 1H), 8.08 (d, J = 5.88Hz, 1H), 7.76-7.97 (m, 1H), 7.43 (d ,J＝7.88Hz,1H),6.52(d,J＝8.00Hz,1H),6.21-6.41(m,1H),4.61-4.75 (m,2H),4.45-4.57(m,1H),4.24(t,J＝6.00Hz,1H),3.82(t,J＝7.07Hz,1H),3.68-3.78(m,1H),3.49- 3.64(m,3H),3.25-3.34(m,1H),3.14-3.22(m,1H),3.04(dt,J＝13.20,8.79Hz,1H),2.72-2.82(m,1H),2.41- 2.54(m,1H),2.24-2.35(m,1H),2.11-2.21(m,1H),2.06(m,1H),1.88-2.01(m,3H),1.48-1.58(m,3H), 1.46(d,J=6.13Hz,3H), 1.36(t,J=7.44Hz,6H). SFC (column: Chiralpak IC-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~50%, 4min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi), Rt=1.411min, ee: 100%.
实施例21、22
Examples 21 and 22
步骤1step 1
将1,4-二氧六环(4mL)和化合物9c(80mg,210.58μmol)加入反应瓶中，搅拌；然后加入碳酸铯(205.83mg,631.74μmol)，化合物1h-2(38.11mg,168.46μmol)，氮气置换，加入BrettPhos Pd G3(57.27mg,63.17μmol)加入，升温至100℃，反应5小时。向反应体系中加入饱和氯化铵溶液(15mL)和乙酸乙酯(15mL)，分液，水相用乙酸乙酯(15mL×2次)萃取，合并有机相，饱和食盐水(15mL)洗涤，无水硫酸钠干燥，过滤，滤液在45℃下进行浓缩得到粗品，粗品通过柱层析纯化(二氯甲烷：甲醇＝40:1～20:1)，然后经SFC拆分(色谱柱：DAICEL CHIRALCEL OD(250mm×30mm，10μm)；流动相：A(超临界CO2)和B(乙醇，含0.1％氨水)；梯度：B％＝50％-50％，12min)得化合物21和化合物22。Add 1,4-dioxane (4mL) and compound 9c (80mg, 210.58μmol) into the reaction bottle and stir; then add cesium carbonate (205.83mg, 631.74μmol), compound 1h-2 (38.11mg, 168.46μmol) ), replace with nitrogen, add BrettPhos Pd G3 (57.27 mg, 63.17 μmol), raise the temperature to 100°C, and react for 5 hours. Add saturated ammonium chloride solution (15mL) and ethyl acetate (15mL) to the reaction system, separate the layers, extract the aqueous phase with ethyl acetate (15mL×2 times), combine the organic phases, and wash with saturated brine (15mL). Dry with anhydrous sodium sulfate and filter. The filtrate is concentrated at 45°C to obtain a crude product. The crude product is purified by column chromatography (dichloromethane: methanol = 40:1 ~ 20:1), and then separated by SFC (chromatography column: DAICEL CHIRALCEL OD (250mm×30mm, 10μm); mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% ammonia); gradient: B%=50%-50%, 12min) to obtain compound 21 and compound 22 .
化合物21：MS m/z:570.0[M+1]+；1H NMR(400MHz,CDCl3)δppm 9.01(s,1H),8.43(s,1H),8.12(d,J＝5.2Hz,1H),8.04-8.02(m,1H),7.40-7.34(m,1H),6.22(d,J＝5.6Hz,1H),4.83-4.69(m,1H),4.64-4.54(m,2H),4.50-4.41(m,2H),4.27-4.17(m,1H),3.79-3.69(m,1H),3.61-3.50(m,2H),3.48-3.35(m,2H),3.26-3.17(m,2H),3.09-3.01(m,1H),2.53-2.46(m,1H),2.33-2.11(m,2H),1.98-1.86(m,4H),1.51(d,J＝13.2,3H),1.38(d,J＝7.2Hz,6H)。SFC(色谱柱：Chiralcel OD-3，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(EtOH，含0.1％异丙胺)；梯度：B％＝5～50％，3min；流速:3.4mL/min；波长:220nm；压力:1800psi，Rt＝1.634min，ee：100％。Compound 21: MS m/z: 570.0[M+1] + ; 1 H NMR (400MHz, CDCl 3 ) δppm 9.01 (s, 1H), 8.43 (s, 1H), 8.12 (d, J = 5.2Hz, 1H ),8.04-8.02(m,1H),7.40-7.34(m,1H),6.22(d,J＝5.6Hz,1H),4.83-4.69(m,1H),4.64-4.54(m,2H), 4.50-4.41(m,2H),4.27-4.17(m,1H),3.79-3.69(m,1H),3.61-3.50(m,2H),3.48-3.35(m,2H),3.26-3.17(m ,2H),3.09-3.01(m,1H),2.53-2.46(m,1H),2.33-2.11(m,2H),1.98-1.86(m,4H),1.51(d,J＝13.2,3H) ,1.38(d,J=7.2Hz,6H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (supercritical CO 2 ) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 ~ 50% , 3min; flow rate: 3.4mL/min; wavelength: 220nm; pressure: 1800psi, Rt=1.634min, ee: 100%.
化合物22：MS m/z:570.0[M+1]+；1H NMR(400MHz,CDCl3)δppm 9.02(s,1H),8.44(s,1H),8.13(d,J＝6.0Hz,1H),8.03-7.91(m,1H),7.47-7.40(m,1H),6.23(d,J＝6.0Hz,1H),4.91-4.73(m,1H),4.71-4.53(m,2H),4.49-4.39(m,2H),4.35-4.19(m,1H),3.77-3.71(m,1H),3.60-3.51(m,2H),3.47-3.34(m,2H),3.26-3.18 (m,2H),3.09-3.02(m,1H),2.52-2.49(m,1H),2.32-2.13(m,2H),1.97-1.86(m,4H),1.51(d,J＝22.0,3H),1.38-1.36(d,J＝7.2Hz,6H)。SFC(色谱柱：Chiralcel OD-3，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(EtOH，含0.1％异丙胺)；梯度：B％＝5～50％，3min；流速:3.4mL/min；波长:220nm；压力:1800psi，Rt＝1.818min，ee：99.66％。Compound 22: MS m/z: 570.0[M+1] + ; 1 H NMR (400MHz, CDCl 3 ) δppm 9.02 (s, 1H), 8.44 (s, 1H), 8.13 (d, J = 6.0Hz, 1H ),8.03-7.91(m,1H),7.47-7.40(m,1H),6.23(d,J＝6.0Hz,1H),4.91-4.73(m,1H),4.71-4.53(m,2H), 4.49-4.39(m,2H),4.35-4.19(m,1H),3.77-3.71(m,1H),3.60-3.51(m,2H),3.47-3.34(m,2H),3.26-3.18 (m,2H),3.09-3.02(m,1H),2.52-2.49(m,1H),2.32-2.13(m,2H),1.97-1.86(m,4H),1.51(d,J＝22.0, 3H), 1.38-1.36 (d, J=7.2Hz, 6H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (supercritical CO 2 ) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 ~ 50% , 3min; flow rate: 3.4mL/min; wavelength: 220nm; pressure: 1800psi, Rt=1.818min, ee: 99.66%.
实施例23
Example 23
步骤1step 1
在反应瓶中加入化合物15g(60mg,210.84μmol)，化合物23a(41.30mg,253.01μmol)，二氧六环(2mL)，碳酸铯(206.09mg,632.51μmol)，氮气置换三次加入Xantphos Pd G4(40.58mg,42.17μmol)，100℃搅拌2小时。反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物23b，MS m/z:367.3[M+1]+。Add compound 15g (60mg, 210.84μmol), compound 23a (41.30mg, 253.01μmol), dioxane (2mL), cesium carbonate (206.09mg, 632.51μmol) into the reaction bottle, replace with nitrogen three times and add Xantphos Pd G4 ( 40.58 mg, 42.17 μmol), stir at 100°C for 2 hours. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane: methanol = 10:0-10:1) to obtain compound 23b, MS m/z: 367.3 [M+1] + .
步骤2Step 2
在反应瓶中加入化合物23b(40mg,109.02μmol)，化合物23f(29.60mg,130.82μmol)，二氧六环(1mL)，碳酸铯(106.56mg,327.06μmol)，氮气置换三次加入BrettPhos Pd G3(19.77mg,21.80μmol)，100℃搅拌4小时。将反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到产物粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物23。MS m/z:557.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.29(s,1H),8.64(s,1H),8.09(d,J＝5.63Hz,1H),7.46(d,J＝7.88Hz,1H),7.36(s,1H),6.96(d,J＝7.88Hz,1H),6.20(d,J＝5.63Hz,1H),4.73-4.96(m,1H),4.59(dt,J＝13.76,6.63Hz,1H),4.25-4.38(m,1H),3.77-3.95(m,1H),3.56-3.68(m,5H),3.53(s,3H),3.00-3.12(m,1H),2.94(s,3H),2.81-2.92(m,2H),2.35(m,2H),2.18-2.29(m,2H),2.05(s,1H),1.81-1.91(m,1H),1.39(dd,J＝6.82,2.44Hz,6H)。Add compound 23b (40 mg, 109.02 μmol), compound 23f (29.60 mg, 130.82 μmol), dioxane (1 mL), and cesium carbonate (106.56 mg, 327.06 μmol) into the reaction bottle. After nitrogen replacement three times, add BrettPhos Pd G3 ( 19.77 mg, 21.80 μmol), stir at 100°C for 4 hours. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 23. MS m/z:557.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.29 (s, 1H), 8.64 (s, 1H), 8.09 (d, J = 5.63Hz, 1H), 7.46 (d, J = 7.88Hz, 1H), 7.36 ( s,1H),6.96(d,J＝7.88Hz,1H),6.20(d,J＝5.63Hz,1H),4.73-4.96(m,1H),4.59(dt,J＝13.76,6.63Hz,1H ),4.25-4.38(m,1H),3.77-3.95(m,1H),3.56-3.68(m,5H),3.53(s,3H),3.00-3.12(m,1H),2.94(s,3H ),2.81-2.92(m,2H),2.35(m,2H),2.18-2.29(m,2H),2.05(s,1H),1.81-1.91(m,1H),1.39(dd,J＝6.82 ,2.44Hz,6H).
实施例24、25
Examples 24 and 25
步骤1step 1
将1,4-二氧六环(6mL)，化合物15g(0.2g,702.79μmol)，化合物9b(178.55mg,843.35μmol,盐酸盐)加入长管中，开始搅拌；氮气置换后，依次将碳酸铯(915.94mg,2.81mmol)，Xantphos Pd G4(67.64mg,70.28μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入10mL饱和氯化铵溶液以及10mL乙酸乙酯后，分液；得到的水相用乙酸乙酯10mL萃取后，合并有机相，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝5:1～1:3)得到化合物24a。 MS m/z:379.0[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.12(s,1H),7.85(s,1H),7.48(d,J＝8.0Hz,1H),6.53(d,J＝8.0Hz,1H),4.45-4.41(m,2H),4.16-3.98(m,2H),3.50-3.42(m,2H),3.24-3.01(m,3H),2.51-2.43(m,1H),2.33-2.11(m,2H),1.96-1.86(m,1H),1.34(d,J＝7.2Hz,6H)。Add 1,4-dioxane (6mL), compound 15g (0.2g, 702.79μmol), and compound 9b (178.55mg, 843.35μmol, hydrochloride) into a long tube and start stirring; after nitrogen replacement, add the Cesium carbonate (915.94 mg, 2.81 mmol) and Xantphos Pd G4 (67.64 mg, 70.28 μmol) were added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. After adding 10 mL of saturated ammonium chloride solution and 10 mL of ethyl acetate to the reaction system, separate the liquids; extract the obtained aqueous phase with 10 mL of ethyl acetate, combine the organic phases, add anhydrous sodium sulfate, dry, filter, and the filtrate is at 45 The crude product was obtained by concentrating under reduced pressure at ℃. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1~1:3) to obtain compound 24a. MS m/z:379.0[M+1] + . 1 H NMR (400MHz, CDCl3) δ = 9.12 (s, 1H), 7.85 (s, 1H), 7.48 (d, J = 8.0Hz, 1H), 6.53 (d, J = 8.0Hz, 1H), 4.45- 4.41(m,2H),4.16-3.98(m,2H),3.50-3.42(m,2H),3.24-3.01(m,3H),2.51-2.43(m,1H),2.33-2.11(m,2H ), 1.96-1.86 (m, 1H), 1.34 (d, J = 7.2Hz, 6H).
步骤2Step 2
将1,4二氧六环(2mL)，化合物24a(20mg,52.78μmol)，化合物23f(10.56mg,47.50μmol)加入长管中，开始搅拌；氮气置换后，将碳酸铯(42.99mg,131.96μmol)，BrettPhos Pd G3(9.57mg,10.56μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入10mL饱和氯化铵溶液以及10mL乙酸乙酯后，分液；水相用乙酸乙酯10mL萃取后，合并有机相，加入无水硫酸钠干燥后，过滤，滤液在45℃下减压浓缩得到粗品。粗品经薄层色谱法纯化(展开剂为乙酸乙酯：甲醇＝15:1)后，制备SFC拆分(色谱柱：Phenomenex-Cellulose-2(250mm×30mm，10μm)；流动相：A(超临界CO2)和B(乙醇，含0.1％氨水)；梯度：B％＝55％-55％，30min)得到化合物24和化合物25。Add 1,4 dioxane (2mL), compound 24a (20mg, 52.78μmol), and compound 23f (10.56mg, 47.50μmol) into the long tube and start stirring; after nitrogen replacement, add cesium carbonate (42.99mg, 131.96 μmol), BrettPhos Pd G3 (9.57 mg, 10.56 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 3 hours. Add 10 mL of saturated ammonium chloride solution and 10 mL of ethyl acetate to the reaction system, and separate the liquids; extract the aqueous phase with 10 mL of ethyl acetate, combine the organic phases, add anhydrous sodium sulfate, dry, and filter, and the filtrate is dried at 45°C. Concentrate under reduced pressure to obtain crude product. After the crude product was purified by thin layer chromatography (the developing solvent is ethyl acetate: methanol = 15:1), the crude product was separated by SFC (chromatographic column: Phenomenex-Cellulose-2 (250mm×30mm, 10μm); mobile phase: A (ultra Critical CO 2 ) and B (ethanol, containing 0.1% ammonia); gradient: B% = 55%-55%, 30min) to obtain compound 24 and compound 25.
化合物24：MS m/z:569.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.06(s,1H),8.56(s,1H),8.08(d,J＝6.0Hz,1H),7.42-7.39(m,2H),6.41(d,J＝8.0Hz,1H),6.21(d,J＝5.6Hz,1H),4.91-4.77(m,1H),4.63-4.56(m,1H),4.46-4.30(m,3H),4.17-4.14(m,1H),3.98-3.95(m,1H),3.87-3.77(m,1H),3.66-3.56(m,3H),3.54-3.44(m,4H),3.24-3.01(m,3H),2.51-2.44(m,1H),2.33-2.13(m,2H),2.09-2.02(m,1H),1.97-1.84(m,2H),1.38-1.36(m,6H)。SFC(色谱柱：Lux Cellulose-2，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝1.648min，ee：100％。Compound 24: MS m/z: 569.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.06 (s, 1H), 8.56 (s, 1H), 8.08 (d, J = 6.0Hz, 1H), 7.42-7.39 (m, 2H), 6.41 (d, J＝8.0Hz,1H),6.21(d,J＝5.6Hz,1H),4.91-4.77(m,1H),4.63-4.56(m,1H),4.46-4.30(m,3H),4.17-4.14 (m,1H),3.98-3.95(m,1H),3.87-3.77(m,1H),3.66-3.56(m,3H),3.54-3.44(m,4H),3.24-3.01(m,3H) ,2.51-2.44(m,1H),2.33-2.13(m,2H),2.09-2.02(m,1H),1.97-1.84(m,2H),1.38-1.36(m,6H). SFC (column: Lux Cellulose-2, 3μm, 0.46cm id×5cm L; mobile phase: A (supercritical CO2) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~50%, 4min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=1.648min, ee: 100%.
化合物25：MS m/z:569.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.06(s,1H),8.56(s,1H),8.08(d,J＝5.6Hz,1H),7.42-7.38(m,2H),6.41(d,J＝8.0Hz,1H),6.21(d,J＝5.6Hz,1H),4.91-4.77(m,1H),4.63-4.56(m,1H),4.46-4.30(m,3H),4.17-4.14(m,1H),3.98-3.95(m,1H),3.87-3.77(m,1H),3.66-3.54(m,3H),3.52(s,3H),3.51-3.44(m,1H),3.21-3.01(m,3H),2.52-2.44(m,1H),2.31-2.02(m,3H),1.97-1.83(m,2H),1.38-1.36(m,6H)。SFC(色谱柱：Lux Cellulose-2，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝2.269min，ee：100％。Compound 25: MS m/z: 569.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.06 (s, 1H), 8.56 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.42-7.38 (m, 2H), 6.41 (d, J＝8.0Hz,1H),6.21(d,J＝5.6Hz,1H),4.91-4.77(m,1H),4.63-4.56(m,1H),4.46-4.30(m,3H),4.17-4.14 (m,1H),3.98-3.95(m,1H),3.87-3.77(m,1H),3.66-3.54(m,3H),3.52(s,3H),3.51-3.44(m,1H),3.21 -3.01(m,3H),2.52-2.44(m,1H),2.31-2.02(m,3H),1.97-1.83(m,2H),1.38-1.36(m,6H). SFC (column: Lux Cellulose-2, 3μm, 0.46cm id×5cm L; mobile phase: A (supercritical CO2) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~50%, 4min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=2.269min, ee: 100%.
实施例26、27
Examples 26 and 27
步骤1step 1
在反应瓶中加入化合物19e(50mg,221.50μmol,盐酸盐)，化合物15g(63.03mg,221.50μmol)，二氧六环(2mL)，碳酸铯(216.50mg,664.49μmol)，氮气置换三次加入BrettPhos Pd G3(31.97mg,33.22μmol)，100℃搅拌8小时。反应液缓慢加入到水溶液(10mL)中淬灭，然后用乙酸乙酯萃取(30mL×2)，收集有机相，有机相用无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)得到化合物26a。MS m/z:393.2[M+1]+。Add compound 19e (50 mg, 221.50 μmol, hydrochloride), compound 15g (63.03 mg, 221.50 μmol), dioxane (2 mL), and cesium carbonate (216.50 mg, 664.49 μmol) into the reaction bottle, and replace with nitrogen three times. BrettPhos Pd G3 (31.97 mg, 33.22 μmol), stirred at 100°C for 8 hours. The reaction solution was slowly added to the aqueous solution (10 mL) to quench, and then extracted with ethyl acetate (30 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 26a. MS m/z:393.2[M+1] + .
步骤2Step 2
将二氧六环(5mL)，化合物26a(45mg,114.52μmol)，化合物23f(22.91mg,103.07μmol)加入长管中，开始搅拌；氮气置换，加入碳酸铯(93.28mg,286.30μmol)，BrettPhos Pd G3(20.76mg,22.90μmol)，升温 至100℃，反应3小时。加入饱和氯化铵溶液(10mL)和乙酸乙酯(10mL)，分液；水相用乙酸乙酯(10mL)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。薄层色谱法纯化(二氯甲烷：甲醇＝10:1)，然后制备SFC拆分(色谱柱：DAICEL CHIRALPAK IC(250mm×30mm,10μm)；流动相：A(超临界CO2)和B(异丙醇，含0.1％氨水)；梯度：B％＝64％-64％，20min)得到化合物26和化合物27。化合物26：MS m/z:583.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.08(s,1H),8.57(s,1H),8.08(d,J＝5.63Hz,1H),7.41(d,J＝8.00Hz,2H),6.53(d,J＝7.88Hz,1H),6.20(dd,J＝5.50Hz,1H),4.72-5.00(m,1H),4.60(t,J＝7.57Hz,2H),4.41(t,J＝6.00Hz,1H),4.26-4.35(m,1H),3.76-3.92(m,1H),3.55-3.69(m,4H),3.48-3.55(m,3H),3.26-3.36(m,1H),3.19(ddd,J＝12.88,8.44,4.06Hz,1H),3.03(ddd,J＝13.29,9.47,8.13Hz,1H),2.78-2.91(m,1H),2.21-2.39(m,2H),2.03-2.17(m,2H),1.81-1.94(m,2H),1.54(d,J＝6.13Hz,3H),1.37(t,J＝6.32Hz,6H)分析SFC(色谱柱：Chiralpak IC-3,50×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi)，Rt＝1.245min，ee：100％。Add dioxane (5mL), compound 26a (45mg, 114.52μmol), and compound 23f (22.91mg, 103.07μmol) into a long tube, start stirring; replace with nitrogen, add cesium carbonate (93.28mg, 286.30μmol), BrettPhos Pd G3 (20.76mg, 22.90μmol), heating to 100°C and react for 3 hours. Add saturated ammonium chloride solution (10 mL) and ethyl acetate (10 mL), separate the layers; extract the aqueous phase with ethyl acetate (10 mL), combine the organic phases, dry over anhydrous sodium sulfate, filter, and reduce the filtrate at 45°C. Pressure concentration. Thin layer chromatography purification (dichloromethane: methanol = 10:1), and then preparative SFC separation (chromatographic column: DAICEL CHIRALPAK IC (250mm×30mm, 10μm); mobile phase: A (supercritical CO 2 ) and B ( Isopropyl alcohol, containing 0.1% ammonia); gradient: B%=64%-64%, 20min) to obtain compound 26 and compound 27. Compound 26: MS m/z: 583.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.08 (s, 1H), 8.57 (s, 1H), 8.08 (d, J = 5.63Hz, 1H), 7.41 (d, J = 8.00Hz, 2H), 6.53 ( d,J＝7.88Hz,1H),6.20(dd,J＝5.50Hz,1H),4.72-5.00(m,1H),4.60(t,J＝7.57Hz,2H),4.41(t,J＝6.00 Hz,1H),4.26-4.35(m,1H),3.76-3.92(m,1H),3.55-3.69(m,4H),3.48-3.55(m,3H),3.26-3.36(m,1H), 3.19(ddd,J＝12.88,8.44,4.06Hz,1H),3.03(ddd,J＝13.29,9.47,8.13Hz,1H),2.78-2.91(m,1H),2.21-2.39(m,2H), 2.03-2.17(m,2H),1.81-1.94(m,2H),1.54(d,J＝6.13Hz,3H),1.37(t,J＝6.32Hz,6H) Analysis SFC (Column: Chiralpak IC- 3,50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~50%, 4min; flow rate: 4.0mL/min ; Wavelength: 220nm; Pressure: 1800psi), Rt=1.245min, ee: 100%.
化合物27：MS m/z:583.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.08(s,1H),8.57(s,1H),8.08(d,J＝5.63Hz,1H),7.40(d,J＝7.88Hz,2H),6.50(d,J＝7.88Hz,1H),6.20(dd,J＝4.88Hz,1H),4.75-4.93(m,1H),4.69(t,J＝7.69Hz,1H),4.58(dt,J＝13.63,7.07Hz,1H),4.27-4.35(m,1H),4.23(t,J＝6.07Hz,1H),3.77-3.92(m,2H),3.55-3.70(m,3H),3.53(s,3H),3.30(dd,J＝7.38Hz,1H),3.17(s,1H),3.05(m,1H),2.70-2.84(m,1H),2.47(m,1H),2.23-2.34(m,1H),2.12-2.21(m,1H),2.03-2.11(m,1H),1.79-1.97(m,2H),1.46(d,J＝6.13Hz,3H),1.37(t,J＝6.25Hz,6H)。分析SFC(色谱柱：Chiralpak IC-3,50×4.6mm I.D.,3μm；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝2.025min，ee：98.52％。Compound 27: MS m/z: 583.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.08 (s, 1H), 8.57 (s, 1H), 8.08 (d, J = 5.63Hz, 1H), 7.40 (d, J = 7.88Hz, 2H), 6.50 ( d,J＝7.88Hz,1H),6.20(dd,J＝4.88Hz,1H),4.75-4.93(m,1H),4.69(t,J＝7.69Hz,1H),4.58(dt,J＝13.63 ,7.07Hz,1H),4.27-4.35(m,1H),4.23(t,J＝6.07Hz,1H),3.77-3.92(m,2H),3.55-3.70(m,3H),3.53(s, 3H),3.30(dd,J＝7.38Hz,1H),3.17(s,1H),3.05(m,1H),2.70-2.84(m,1H),2.47(m,1H),2.23-2.34(m ,1H),2.12-2.21(m,1H),2.03-2.11(m,1H),1.79-1.97(m,2H),1.46(d,J＝6.13Hz,3H),1.37(t,J＝6.25 Hz, 6H). Analytical SFC (chromatographic column: Chiralpak IC-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~50% , 4min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=2.025min, ee: 98.52%.
实施例28
Example 28
步骤1step 1
将异丙醇(10mL)，化合物14e(200mg,522.54μmol)，化合物28h(102.36mg,盐酸盐)加入封管中，搅拌；然后将N,N-二异丙基乙胺(236.37mg,1.83mmol,318.56μL,3.5eq)加入其中，升温至100℃，反应1小时。反应液降至室温20℃后，向反应体系中加入30mL水后，加入乙酸乙酯萃取(15mL×2)；得到的有机相用15mL饱和食盐水洗涤一次后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～20:80)，得到化合物28a。MS m/z:396.0[M+1]+。1H NMR(400MHz,CDCl3)δppm 8.84(s,1H),7.32(s,1H),6.21(s,1H),5.04-4.96(m,2H),4.90(t,J＝8.4,1H),4.80-4.73(m,2H),4.69-4.63(m,1H),4.16-4.12(m,1H),3.44-3.32(m,2H),3.10- 3.05(m,1H),2.98(s,3H),1.83(s,3H),1.66(d,J＝6.0,3H)。Add isopropyl alcohol (10 mL), compound 14e (200 mg, 522.54 μmol), and compound 28h (102.36 mg, hydrochloride) into the sealed tube, stir; then add N, N-diisopropylethylamine (236.37 mg, 1.83mmol, 318.56μL, 3.5eq) was added, the temperature was raised to 100°C, and the reaction was carried out for 1 hour. After the reaction solution cools down to room temperature 20°C, add 30 mL of water to the reaction system, add ethyl acetate for extraction (15 mL × 2); wash the obtained organic phase once with 15 mL of saturated brine, add anhydrous sodium sulfate, dry, and filter. , the filtrate was concentrated under reduced pressure at 45°C to obtain crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0~20:80) to obtain compound 28a. MS m/z:396.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 8.84 (s, 1H), 7.32 (s, 1H), 6.21 (s, 1H), 5.04-4.96 (m, 2H), 4.90 (t, J = 8.4, 1H) ,4.80-4.73(m,2H),4.69-4.63(m,1H),4.16-4.12(m,1H),3.44-3.32(m,2H),3.10- 3.05(m,1H),2.98(s,3H),1.83(s,3H),1.66(d,J=6.0,3H).
步骤2Step 2
将二氯甲烷(20mL)，化合物28a(190mg,479.92μmol)加入50mL反应瓶中，开始搅拌；然后将温度降至0～5℃后，将N-碘代丁二酰亚胺(107.97mg,479.92μmol)溶于0.5mL无水二氯甲烷中后，滴加到其中，反应0.5小时。向反应体系中加入20mL水后，搅拌5min后，分液；水相用10mL二氯甲烷萃取后，合并有机相加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～20:80)，得到化合物28b。MS m/z:521.9[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.73(s,1H),7.67(s,1H),5.12-4.98(m,2H),4.93(t,J＝8.4,1H),4.88-4.79(m,2H),4.69-4.63(m,1H),4.18-4.14(m,1H),3.44-3.33(m,2H),3.14-3.05(m,1H),2.99(s,3H),1.87(s,3H),1.67(d,J＝6.0,3H)。Add dichloromethane (20 mL) and compound 28a (190 mg, 479.92 μmol) into a 50 mL reaction flask and start stirring; then lower the temperature to 0~5°C, add N-iodosuccinimide (107.97 mg, 479.92 μmol) was dissolved in 0.5 mL of anhydrous methylene chloride, and then added dropwise to it and allowed to react for 0.5 hours. After adding 20 mL of water to the reaction system, stir for 5 minutes and then separate the liquids; extract the aqueous phase with 10 mL of methylene chloride, combine the organic phases and add anhydrous sodium sulfate to dry, then filter, and the filtrate is concentrated under reduced pressure at 45°C. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0~20:80) to obtain compound 28b. MS m/z:521.9[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.73 (s, 1H), 7.67 (s, 1H), 5.12-4.98 (m, 2H), 4.93 (t, J = 8.4, 1H), 4.88-4.79 (m ,2H),4.69-4.63(m,1H),4.18-4.14(m,1H),3.44-3.33(m,2H),3.14-3.05(m,1H),2.99(s,3H),1.87(s ,3H),1.67(d,J=6.0,3H).
步骤3Step 3
将N,N-二甲基甲酰胺(10mL)，化合物28b(100mg,191.64μmol)，加入长管中，搅拌；氮气置换后，然后将甲酸钠(32.58mg,479.11μmol,25.86μL)，四丁基氯化铵(106.52mg,383.29μmol)，乙酸钾(75.23mg,766.58μmol)，醋酸钯(8.61mg,38.33μmol)加入其中，升温至100℃，反应1小时。将反应液降至室温后，向反应体系中加入30mL水，再用乙酸乙酯萃取(15mL×3)；得到的有机相用20mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～10:90)得到化合物28c。MS m/z:396.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.88(s,1H),7.39(s,1H),4.96-4.88(m,1H),4.73-4.67(m,1H),4.33(s,1H),4.20-4.10(m,2H),3.43-3.28(m,2H),3.08-3.00(m,1H),2.97(s,3H),1.63-1.62(m,3H),1.53-1.51(m,6H)。Add N,N-dimethylformamide (10 mL) and compound 28b (100 mg, 191.64 μmol) into a long tube and stir; after nitrogen replacement, add sodium formate (32.58 mg, 479.11 μmol, 25.86 μL), tetrabutyl Ammonium chloride (106.52 mg, 383.29 μmol), potassium acetate (75.23 mg, 766.58 μmol), and palladium acetate (8.61 mg, 38.33 μmol) were added, the temperature was raised to 100°C, and the reaction was carried out for 1 hour. After the reaction solution was brought to room temperature, 30 mL of water was added to the reaction system, and then extracted with ethyl acetate (15 mL × 3); the obtained organic phase was washed with 20 mL of saturated brine, dried by adding anhydrous sodium sulfate, filtered, and the filtrate Concentrate under reduced pressure at 45°C to obtain crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0~10:90) to obtain compound 28c. MS m/z:396.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.88 (s, 1H), 7.39 (s, 1H), 4.96-4.88 (m, 1H), 4.73-4.67 (m, 1H), 4.33 (s, 1H), 4.20-4.10(m,2H),3.43-3.28(m,2H),3.08-3.00(m,1H),2.97(s,3H),1.63-1.62(m,3H),1.53-1.51(m,6H ).
步骤4Step 4
将1,4-二氧六环(2.5mL)，化合物28c(25mg,63.15μmol)，化合物23f(12.86mg,56.83μmol)加入长管中，搅拌；氮气置换后，将BrettPhos Pd G3(11.45mg,12.63μmol)，乙酸钾(18.59mg,189.44μmol)加入其中，升温至90℃，反应2小时。向反应体系中加入10mL水后，加入乙酸乙酯萃取(10mL×2)；有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。薄层层析制备分离(展开剂为乙酸乙酯:甲醇＝10:1)得到化合物28。MS m/z:586.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.83(s,1H),8.11(d,J＝5.2,1H),7.90(s,1H),7.45(s,1H),6.32-6.28(m,1H),4.92-4.76(m,2H),4.73-4.57(m,2H),4.36-4.30(m,3H),4.14-4.07(m,1H),3.79-3.69(m,1H),3.63-3.55(m,2H),3.51(s,3H),3.43-3.28(m,2H),3.06-2.93(m,4H),2.03-1.85(m,2H),1.62(d,J＝6.0,3H),1.57(d,J＝10.4,6H)。Add 1,4-dioxane (2.5mL), compound 28c (25mg, 63.15μmol), and compound 23f (12.86mg, 56.83μmol) into a long tube and stir; after nitrogen replacement, add BrettPhos Pd G3 (11.45mg , 12.63 μmol), potassium acetate (18.59 mg, 189.44 μmol) was added, the temperature was raised to 90°C, and the reaction was carried out for 2 hours. After adding 10 mL of water to the reaction system, add ethyl acetate for extraction (10 mL × 2); wash the organic phase with 10 mL of saturated brine, add anhydrous sodium sulfate, dry, filter, and concentrate the filtrate under reduced pressure at 45°C. Preparative separation by thin layer chromatography (developing solvent: ethyl acetate:methanol=10:1) gave compound 28. MS m/z:586.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.83 (s, 1H), 8.11 (d, J = 5.2, 1H), 7.90 (s, 1H), 7.45 (s, 1H), 6.32-6.28 (m, 1H) ),4.92-4.76(m,2H),4.73-4.57(m,2H),4.36-4.30(m,3H),4.14-4.07(m,1H),3.79-3.69(m,1H),3.63-3.55 (m,2H),3.51(s,3H),3.43-3.28(m,2H),3.06-2.93(m,4H),2.03-1.85(m,2H),1.62(d,J＝6.0,3H) ,1.57(d,J=10.4,6H).
实施例29
Example 29
步骤1 step 1
将甲醇(80mL)，化合物14c(4g,18.99mmol)加入250mL三口瓶中，搅拌；然后将氢化钠(3.04g,75.97mmol,60％纯度)缓慢分批加入其中，升温至50℃，反应3小时。将反应液降至室温，向反应体系中加入150mL饱和氯化铵水溶液，加入乙酸乙酯(100mL×6)萃取；合并有机相，加入200mL饱和食盐水洗涤后，无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品中加入10mL乙酸乙酯以及2mL甲醇，搅拌0.5小时后，过滤，滤饼即化合物29a。MS m/z:211.1[M+1]+。Add methanol (80mL) and compound 14c (4g, 18.99mmol) into a 250mL three-necked flask and stir; then add sodium hydride (3.04g, 75.97mmol, 60% purity) slowly in batches, raise the temperature to 50°C, and react 3 Hour. Lower the reaction solution to room temperature, add 150 mL saturated aqueous ammonium chloride solution to the reaction system, add ethyl acetate (100 mL × 6) for extraction; combine the organic phases, add 200 mL saturated brine, wash, dry over anhydrous sodium sulfate, and filter. The filtrate was concentrated under reduced pressure at 45°C to obtain crude product. Add 10 mL of ethyl acetate and 2 mL of methanol to the crude product, stir for 0.5 hours, and then filter. The filter cake is compound 29a. MS m/z:211.1[M+1] + .
步骤2Step 2
将二氯甲烷(52mL)，化合物29a(1.3g,6.17mmol)加入反应瓶中，搅拌；然后将三乙胺(2.50g,24.69mmol,3.44mL)加入其中，将温度降至0～5℃后，将三氟甲磺酸酐(4.35g,15.43mmol,2.55mL)滴加到其中，反应0.5小时。向反应体系中加入50ml饱和氯化铵溶液以及20mL二氯甲烷后，分液，得到的有机相用50mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品再经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～80:20)，得到粗品化合物29b(约50％纯度)。MS m/z:342.9[M+1]+。Add dichloromethane (52mL) and compound 29a (1.3g, 6.17mmol) into the reaction flask and stir; then add triethylamine (2.50g, 24.69mmol, 3.44mL) and lower the temperature to 0~5°C. After that, trifluoromethanesulfonic anhydride (4.35g, 15.43mmol, 2.55mL) was added dropwise to it, and the reaction was carried out for 0.5 hours. After adding 50 ml of saturated ammonium chloride solution and 20 ml of methylene chloride to the reaction system, the liquids were separated. The obtained organic phase was washed with 50 ml of saturated brine, dried by adding anhydrous sodium sulfate, and filtered. The filtrate was decompressed at 45°C. Concentrate to obtain crude product. The crude product was further purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~80:20) to obtain crude compound 29b (approximately 50% purity). MS m/z:342.9[M+1] + .
步骤3Step 3
将异丙醇(15mL)，化合物29b(1g,1.46mmol,50％纯度)，化合物28h(291.38mg,1.46mmol,盐酸盐)加入反应瓶，搅拌；然后将二异丙基乙胺(660.01mg,5.11mmol,889.50μL)加入其中，升温至100℃，反应0.5小时。向反应体系中加入40mL水后，加入乙酸乙酯萃取(20mL×2)；得到的有机相用20mL饱和食盐水溶液洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～10:90)。得到化合物29c。MS m/z:356.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.86(s,1H),7.33(s,1H),6.16(s,1H),4.92(t,J＝8.0,1H),4.71-4.68(m,1H),4.17-4.13(m,1H),3.95(s,3H),3.44-3.32(m,2H),3.10-3.05(m,1H),2.98(s,3H),1.67(d,J＝6.0,3H)。Add isopropyl alcohol (15mL), compound 29b (1g, 1.46mmol, 50% purity), compound 28h (291.38mg, 1.46mmol, hydrochloride) into the reaction flask, stir; then diisopropylethylamine (660.01 mg, 5.11 mmol, 889.50 μL) was added, the temperature was raised to 100°C, and the reaction was carried out for 0.5 hours. After adding 40mL of water to the reaction system, add ethyl acetate for extraction (20mL×2); wash the obtained organic phase with 20mL of saturated saline solution, add anhydrous sodium sulfate, dry, filter, and concentrate the filtrate under reduced pressure at 45°C. Get crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~10:90). Compound 29c was obtained. MS m/z:356.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.86 (s, 1H), 7.33 (s, 1H), 6.16 (s, 1H), 4.92 (t, J = 8.0, 1H), 4.71-4.68 (m, 1H) ),4.17-4.13(m,1H),3.95(s,3H),3.44-3.32(m,2H),3.10-3.05(m,1H),2.98(s,3H),1.67(d,J＝6.0 ,3H).
步骤4Step 4
将二氯甲烷(20mL)，化合物29c(350mg,983.59μmol)加入烧瓶中，搅拌；然后将温度降至0～5℃后，将N-碘代丁二酰亚胺(221.29mg,983.59μmol)加入其中，反应0.5小时。向反应体系中加入20mL水后，分液；得到的有机相用20mL饱和食盐水溶液洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～10:90)，得到化合物29d。MS m/z:482.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.74(s,1H),7.65(s,1H),4.96-4.91(m,1H),4.72-4.66(m,1H),4.36(d,J＝8.8and 6.0,1H),4.02(s,3H),3.45-3.34(m,2H),3.16-3.06(m,1H),2.99(s,3H),1.68(d,J＝6.4,3H)。Add dichloromethane (20 mL) and compound 29c (350 mg, 983.59 μmol) into the flask and stir; then lower the temperature to 0~5°C, add N-iodosuccinimide (221.29 mg, 983.59 μmol) Add it and react for 0.5 hours. After adding 20 mL of water to the reaction system, the layers were separated; the obtained organic phase was washed with 20 mL of saturated saline solution, dried by adding anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~10:90) to obtain compound 29d. MS m/z:482.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.74 (s, 1H), 7.65 (s, 1H), 4.96-4.91 (m, 1H), 4.72-4.66 (m, 1H), 4.36 (d, J = 8.8 and 6.0,1H),4.02(s,3H),3.45-3.34(m,2H),3.16-3.06(m,1H),2.99(s,3H),1.68(d,J＝6.4,3H).
步骤5Step 5
将1,4-二氧六环(20mL)，水(4mL)，化合物29d(0.26g,539.72μmol)，异丙烯基硼酸嚬哪醇酯(136.04mg,809.57μmol)加入反应瓶中，搅拌；氮气置换后，依次将碳酸钾(186.49mg,1.35mmol)，1,1-双(二苯基磷)二茂铁氯化钯二氯甲烷络合物(88.15mg,107.94μmol)加入其中，升温至80℃，反应2小时。将反应体系降至室温后，加入30mL饱和氯化铵溶液，然后加入乙酸乙酯萃取(15mL×2)萃取，得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝1:1～1:2.5)得到化合物29e。MS m/z:396.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.88(s,1H),7.55(s,1H),5.48-4.90(m,2H),4.71-4.67(m,1H),4.15-4.11(m,1H),4.00-3.95(m,4H),3.43-3.31(m,2H),3.14-3.06(m,1H),2.98(s,3H),2.04(s,3H),1.70-1.68(m,3H)。Add 1,4-dioxane (20 mL), water (4 mL), compound 29d (0.26 g, 539.72 μmol), isopropenyl borate zonacol ester (136.04 mg, 809.57 μmol) into the reaction bottle, and stir; After nitrogen replacement, potassium carbonate (186.49 mg, 1.35 mmol) and 1,1-bis(diphenylphosphorus)ferrocene palladium chloride dichloromethane complex (88.15 mg, 107.94 μmol) were added successively, and the temperature was raised. to 80°C and react for 2 hours. After the reaction system was brought to room temperature, 30 mL of saturated ammonium chloride solution was added, and then ethyl acetate was added for extraction (15 mL × 2). The obtained organic phase was washed with 15 mL of saturated brine, dried by adding anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure at 45°C to obtain crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:1~1:2.5) to obtain compound 29e. MS m/z:396.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.88 (s, 1H), 7.55 (s, 1H), 5.48-4.90 (m, 2H), 4.71-4.67 (m, 1H), 4.15-4.11 (m, 1H) ),4.00-3.95(m,4H),3.43-3.31(m,2H),3.14-3.06(m,1H),2.98(s,3H),2.04(s,3H),1.70-1.68(m,3H ).
步骤6Step 6
将甲醇(12mL)，化合物29e(150mg,378.88μmol)加入50ml单口瓶中，搅拌；然后将二氧化铂(43.02mg,189.44μmol)加入其中，氢气15psi，室温20℃，反应5小时。反应液直接过滤后，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～15:85)得到化合物29f。MS m/z:398.1 [M+1]+。Add methanol (12 mL) and compound 29e (150 mg, 378.88 μmol) into a 50 ml single-neck bottle and stir; then add platinum dioxide (43.02 mg, 189.44 μmol), hydrogen 15 psi, room temperature 20°C, and react for 5 hours. After the reaction solution was directly filtered, the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~15:85) to obtain compound 29f. MS m/z:398.1 [M+1] + .
步骤7Step 7
将1,4-二氧六环(3mL)，化合物29f(30mg,75.39μmol)，化合物23f(13.65mg,60.31μmol)加入长管中，搅拌；氮气置换后，将乙酸钾(22.20mg,226.18μmol)，BrettPhos Pd G3(13.67mg,15.08μmol)加入其中，升温至90℃，反应2小时。向反应体系中加入15mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相加入15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～15:85)得到粗品，粗品经制备高效液相色谱分离(色谱柱:Waters Xbridge BEH C18 100×30mm×10μm；流动相:A(乙腈)和B(水，含0.05％碳酸氢铵)，梯度：B％:40％-65％)得到化合物29。MS m/z:588.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.84(s,1H),8.40(s,1H),8.09-8.07(m,1H),7.36(s,1H),6.16(d,J＝5.6,1H),4.92-4.78(m,2H),4.70-4.61(m,2H),4.41-4.37(m,1H),4.06-3.97(m,4H),3.76-3.45(m,7H),3.43-3.28(m,2H),3.09-3.02(m,1H),2.97(s,3H),2.08-1.99(m,1H),1.89-1.86(m,1H),1.66-1.65(m,3H),1.37(dd,J＝10.4,7.2,6H)。Add 1,4-dioxane (3mL), compound 29f (30mg, 75.39μmol), and compound 23f (13.65mg, 60.31μmol) into a long tube and stir; after nitrogen replacement, add potassium acetate (22.20mg, 226.18 μmol), BrettPhos Pd G3 (13.67 mg, 15.08 μmol) was added, the temperature was raised to 90°C, and the reaction was carried out for 2 hours. After adding 15 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:0~15:85), and the crude product was separated by preparative high performance liquid chromatography (chromatographic column: Waters Xbridge BEH C18 100×30mm×10μm; mobile phase: A (acetonitrile) and B (water with 0.05% ammonium bicarbonate), gradient: B%: 40%-65%) gave compound 29. MS m/z:588.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.84 (s, 1H), 8.40 (s, 1H), 8.09-8.07 (m, 1H), 7.36 (s, 1H), 6.16 (d, J = 5.6, 1H ),4.92-4.78(m,2H),4.70-4.61(m,2H),4.41-4.37(m,1H),4.06-3.97(m,4H),3.76-3.45(m,7H),3.43-3.28 (m,2H),3.09-3.02(m,1H),2.97(s,3H),2.08-1.99(m,1H),1.89-1.86(m,1H),1.66-1.65(m,3H),1.37 (dd,J=10.4,7.2,6H).
实施例30
Example 30
步骤1step 1
将锌粉(2.31g,35.32mmol)与四氢呋喃(25mL)加入250mL三口瓶中后，搅拌，然后依次将1,2-二溴乙烷(497.68mg,2.65mmol,199.87μL)，三甲基氯硅烷(287.81mg,2.65mmol,336.22μL)加入其中，室温18℃搅拌5分钟；然后将化合物30a(5g,17.66mmol)溶于四氢呋喃(25mL)后，缓慢滴加，室温18℃反应1小时，升温至45℃，继续反应1小时。得到化合物30b悬浊液，反应液直接用于下一步。Add zinc powder (2.31g, 35.32mmol) and tetrahydrofuran (25mL) into a 250mL three-necked flask, stir, and then add 1,2-dibromoethane (497.68mg, 2.65mmol, 199.87μL), trimethyl chloride in sequence Silane (287.81mg, 2.65mmol, 336.22μL) was added and stirred at room temperature 18°C for 5 minutes; then compound 30a (5g, 17.66mmol) was dissolved in tetrahydrofuran (25mL), slowly added dropwise, and reacted at room temperature 18°C for 1 hour. Raise the temperature to 45°C and continue the reaction for 1 hour. A suspension of compound 30b was obtained, and the reaction solution was used directly in the next step.
步骤2Step 2
将N,N-二甲基乙酰胺(8mL),化合物15g(0.2g,702.79μmol)，加入长管中开始搅拌；氮气置换后，将碘化亚铜(40.15mg,210.84μmol)，1,1-双(二苯基磷)二茂铁氯化钯(154.27mg,210.84μmol)，升温至80℃，搅拌10min。然后将化合物30b悬浊液滴加入其中，反应2小时。将反应液降至室温后，向反应体系中加入20mL水后，加入乙酸乙酯萃取(10mL×2)，有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～50:50)，得到化合物30c。MS m/z:361.2[M+1]+。Add N,N-dimethylacetamide (8mL) and compound 15g (0.2g, 702.79μmol) into a long tube and start stirring; after nitrogen replacement, add copper iodide (40.15mg, 210.84μmol), 1, 1-Bis(diphenylphosphorus)ferrocene palladium chloride (154.27 mg, 210.84 μmol), heat up to 80°C, and stir for 10 minutes. Then compound 30b suspension was added dropwise, and the reaction was carried out for 2 hours. After the reaction solution was brought to room temperature, 20 mL of water was added to the reaction system, and ethyl acetate was added for extraction (10 mL × 2). The organic phase was washed with 10 mL of saturated brine, dried by adding anhydrous sodium sulfate, and filtered. The crude product was obtained by concentrating under reduced pressure at ℃. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~50:50) to obtain compound 30c. MS m/z:361.2[M+1]+.
步骤3Step 3
将二氯甲烷(10mL)，化合物30c(110mg,304.81μmol)加入拇指瓶中，搅拌；然后将三氟乙酸(44.57mmol,3.30mL)加入其中，室温20℃，反应2小时。反应液在35℃下进行减压浓缩，用15mL乙酸乙酯 溶解后；依次用15mL饱和碳酸氢钠，15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在40℃下进行减压浓缩。得到化合物30d的三氟乙酸盐。MS m/z:261.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.01(s,1H),7.97(s,1H),7.63(d,J＝7.6Hz,1H),7.47(d,J＝7.6Hz,1H),4.86-4.78(m,1H),4.31-4.17(m,4H),3.62-3.55(m,2H),1.39(d,J＝6.8Hz,6H)。Add dichloromethane (10 mL) and compound 30c (110 mg, 304.81 μmol) into the thumb bottle and stir; then add trifluoroacetic acid (44.57 mmol, 3.30 mL) and react at room temperature 20°C for 2 hours. The reaction solution was concentrated under reduced pressure at 35°C, and 15 mL of ethyl acetate was used. After dissolving, wash with 15 mL saturated sodium bicarbonate and 15 mL saturated saline, add anhydrous sodium sulfate, dry, filter, and concentrate the filtrate under reduced pressure at 40°C. The trifluoroacetate salt of compound 30d was obtained. MS m/z:261.1[M+1]+. 1H NMR (400MHz, CDCl3) δ = 9.01 (s, 1H), 7.97 (s, 1H), 7.63 (d, J = 7.6Hz, 1H), 7.47 (d, J = 7.6Hz, 1H), 4.86-4.78 (m,1H),4.31-4.17(m,4H),3.62-3.55(m,2H),1.39(d,J＝6.8Hz,6H).
步骤4Step 4
将二氯甲烷(5mL)，化合物30d的三氟乙酸盐(70mg,268.44μmol)加入拇指瓶中，搅拌；然后将温度降至0～5℃后，依次将三乙胺(54.33mg,536.89μmol,74.73μL)，甲烷磺酰氯(33.83mg,295.29μmol,22.85μL)滴加到其中，升温至室温20℃，反应0.5小时。将反应液倒入20mL冰水中后，加入10mL二氯甲烷后分液，水相用15mL二氯甲烷萃取后，合并有机相，用20mL饱和食盐水洗涤一次后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～40:60)，得到化合物30e。1H NMR(400MHz,CDCl3)δ＝9.02(s,1H),7.99(s,1H),7.64(d,J＝7.6Hz,1H),7.51(d,J＝7.6Hz,1H),4.63-4.55(m,1H),4.44-4.40(m,2H),4.29-4.25(m,2H),3.63-3.56(m,1H),2.94(s,3H),1.40(d,J＝6.8Hz,6H)。Add dichloromethane (5 mL) and the trifluoroacetate of compound 30d (70 mg, 268.44 μmol) into the thumb bottle and stir; then lower the temperature to 0~5°C, then add triethylamine (54.33 mg, 536.89 μmol, 74.73 μL), methane sulfonyl chloride (33.83 mg, 295.29 μmol, 22.85 μL) was added dropwise, and the temperature was raised to room temperature 20°C, and the reaction was carried out for 0.5 hours. Pour the reaction solution into 20 mL of ice water, add 10 mL of dichloromethane and separate the liquids. Extract the aqueous phase with 15 mL of dichloromethane, combine the organic phases, wash once with 20 mL of saturated brine, add anhydrous sodium sulfate, dry and filter. , the filtrate was concentrated under reduced pressure at 45°C to obtain crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~40:60) to obtain compound 30e. 1H NMR (400MHz, CDCl3) δ = 9.02 (s, 1H), 7.99 (s, 1H), 7.64 (d, J = 7.6Hz, 1H), 7.51 (d, J = 7.6Hz, 1H), 4.63-4.55 (m,1H),4.44-4.40(m,2H),4.29-4.25(m,2H),3.63-3.56(m,1H),2.94(s,3H),1.40(d,J＝6.8Hz,6H ).
步骤5Step 5
将1,4-二氧六环(2mL)，化合物30e(20mg,59.02μmol)，化合物23f(13.35mg,59.02μmol)加入长管中，开始搅拌；氮气置换后，依次将乙酸钾(17.38mg,177.07μmol)，BrettPhos Pd G3(10.70mg,11.80μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入15mL水后，加入乙酸乙酯萃取(15mL×2)；得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层硅胶色谱纯化(二氯甲烷：甲醇＝10:1)得到化合物30。MS m/z:529.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.92(s,1H),8.73(s,1H),8.10(d,J＝5.6Hz,1H),7.56(d,J＝7.6Hz,1H),7.34-7.33(m,2H),6.19(d,J＝5.6Hz,1H),4.92-4.78(m,1H),4.63-4.55(m,2H),4.44(d,J＝8.0Hz,2H),4.32-4.25(m,3H),3.89-3.80(m,1H),3.73-3.62(m,3H),3.53(s,3H),2.94(s,3H),2.10-2.03(m,1H),1.91-1.84(m,1H),1.42(dd,J＝6.8and 2.8Hz,6H)。Add 1,4-dioxane (2mL), compound 30e (20mg, 59.02μmol), and compound 23f (13.35mg, 59.02μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate (17.38mg) ,177.07 μmol), BrettPhos Pd G3 (10.70 mg, 11.80 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. After adding 15 mL of water to the reaction system, add ethyl acetate for extraction (15 mL Get crude product. The crude product was purified by thin-layer silica gel chromatography (dichloromethane:methanol=10:1) to obtain compound 30. MS m/z:529.4[M+1]+. 1H NMR (400MHz, CDCl3) δ = 8.92 (s, 1H), 8.73 (s, 1H), 8.10 (d, J = 5.6Hz, 1H), 7.56 (d, J = 7.6Hz, 1H), 7.34-7.33 (m,2H),6.19(d,J＝5.6Hz,1H),4.92-4.78(m,1H),4.63-4.55(m,2H),4.44(d,J＝8.0Hz,2H),4.32- 4.25(m,3H),3.89-3.80(m,1H),3.73-3.62(m,3H),3.53(s,3H),2.94(s,3H),2.10-2.03(m,1H),1.91- 1.84(m,1H),1.42(dd,J=6.8and 2.8Hz,6H).
实施例31、32
Examples 31 and 32
步骤1step 1
在反应瓶中加入化合物4h(80mg,331.79μmol)，化合物19e(82.39mg,364.97μmol,盐酸盐)，异丙醇(4mL)，三乙胺(201.44mg,1.99mmol)，氮气置换三次加入100℃搅拌4小时。反应液进行减压浓缩得到粗品，用快速硅胶柱层析纯化(二氯甲烷：甲醇＝10:0～10:1)得到化合物31a。MS m/z:394.3/396.3[M+1]+。Add compound 4h (80 mg, 331.79 μmol), compound 19e (82.39 mg, 364.97 μmol, hydrochloride), isopropyl alcohol (4 mL), and triethylamine (201.44 mg, 1.99 mmol) into the reaction bottle, and replace with nitrogen three times. Stir at 100°C for 4 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by flash silica gel column chromatography (dichloromethane: methanol = 10:0 to 10:1) to obtain compound 31a. MS m/z:394.3/396.3[M+1] + .
步骤2Step 2
在反应瓶中加入31a(40mg,101.54μmol)，1h-2(22.97mg,101.54μmol)，二氧六环(2mL)，碳酸铯(99.25mg,304.62μmol)氮气置换三次加入BrettPhos Pd G3(18.41mg,20.31μmol)，100℃搅拌2小时。加入水(4mL)，用乙酸乙酯萃取(5mL×3)，合并有机相，用饱和食盐水洗涤(5mL)，无水硫酸钠干燥，过滤，减压浓缩得到粗品，粗品经柱层析(二氯甲烷：甲醇＝10:0～10:1)纯化，然后经制备SFC拆分(色谱柱：DAICEL CHIRALPAK IC(250mm×30mm,10μm)；流动相：A(超临界CO2)和B(异丙醇，含0.1％氨水)；梯度：B％＝42％-42％，15min)得到化合物31和化合物32。Add 31a (40 mg, 101.54 μmol), 1h-2 (22.97 mg, 101.54 μmol), dioxane (2 mL), and cesium carbonate (99.25 mg, 304.62 μmol) to the reaction bottle. Nitrogen replacement was performed three times and BrettPhos Pd G3 (18.41 mg, 20.31 μmol), stir at 100°C for 2 hours. Add water (4 mL), extract with ethyl acetate (5 mL Dichloromethane: methanol = 10:0 ~ 10:1) purification, and then preparative SFC separation (chromatographic column: DAICEL CHIRALPAK IC (250mm×30mm, 10μm); mobile phase: A (supercritical CO 2 ) and B ( Isopropyl alcohol, containing 0.1% ammonia); gradient: B%=42%-42%, 15min) to obtain compound 31 and compound 32.
化合物31：MS m/z:584.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.01(s,1H),8.42(s,1H),8.12(d,J＝5.63Hz,1H),8.05(s,1H),7.37(s,1H),6.21(d,J＝5.63Hz,1H),4.73-4.89(m,2H),4.60(m,1H),4.40-4.52(m,1H),3.85-3.97(m,1H),3.67-3.80(m,1H),3.49-3.62(m,2H),3.33-3.46(m,2H),3.14-3.24(m,1H),3.02(dt,J＝13.07,8.85Hz,1H),2.77-2.88(m,1H),2.35-2.45(m,1H),2.20-2.30(m,1H),2.07-2.17(m,1H),1.91-2.01(m,2H),1.88(m,1H),1.60(m,3H),1.44-1.54(m,4H),1.38(dd,J＝6.75,3.00Hz,6H)。分析SFC(色谱柱：Chiralpak AD-3,50×4.6mm I.D,3μm；流动相：A(超临界CO2)和B(异丙醇，含0.1％异丙胺)；梯度：B％＝50～50％，3min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝0.641min。ee值：100％Compound 31: MS m/z: 584.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.01 (s, 1H), 8.42 (s, 1H), 8.12 (d, J = 5.63Hz, 1H), 8.05 (s, 1H), 7.37 (s, 1H), 6.21(d,J＝5.63Hz,1H),4.73-4.89(m,2H),4.60(m,1H),4.40-4.52(m,1H),3.85-3.97(m,1H),3.67-3.80( m,1H),3.49-3.62(m,2H),3.33-3.46(m,2H),3.14-3.24(m,1H),3.02(dt,J＝13.07,8.85Hz,1H),2.77-2.88( m,1H),2.35-2.45(m,1H),2.20-2.30(m,1H),2.07-2.17(m,1H),1.91-2.01(m,2H),1.88(m,1H),1.60( m,3H),1.44-1.54(m,4H),1.38(dd,J=6.75,3.00Hz,6H). Analytical SFC (chromatographic column: Chiralpak AD-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (isopropanol, containing 0.1% isopropylamine); gradient: B%=50~ 50%, 3min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=0.641min. ee value: 100%
化合物32：MS m/z:584.4[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.02(s,1H),8.43(s,1H),8.12(d,J＝5.25Hz,1H),7.98-8.06(m,1H),7.39(s,1H),6.21(d,J＝5.25Hz,1H),4.87-4.99(m,1H),4.51-4.69(m,2H),4.38-4.48(m,1H),4.06-4.18(m,1H),3.67-3.84(m,1H),3.47-3.64(m,2H),3.34-3.45(m,1H),3.21-3.31(m,1H),3.11-3.20(m,1H),2.95-3.07(m,1H),2.73-2.85(m,1H),2.39-2.52(m,1H),2.25(s,1H),2.08-2.18(m,1H),1.93-1.98(m,2H),1.80-1.89(m,1H),1.58-1.63(m,3H),1.49(s,4H),1.38(m,6H)。SFC(色谱柱：Chiralpak AD-3,50×4.6mm I.D,3μm；流动相：A(超临界CO2)和B(异丙醇，含0.1％异丙胺)；梯度：B％＝50～50％，3min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝1.367min。ee值：99％Compound 32: MS m/z: 584.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.02 (s, 1H), 8.43 (s, 1H), 8.12 (d, J = 5.25Hz, 1H), 7.98-8.06 (m, 1H), 7.39 (s, 1H) ),6.21(d,J＝5.25Hz,1H),4.87-4.99(m,1H),4.51-4.69(m,2H),4.38-4.48(m,1H),4.06-4.18(m,1H), 3.67-3.84(m,1H),3.47-3.64(m,2H),3.34-3.45(m,1H),3.21-3.31(m,1H),3.11-3.20(m,1H),2.95-3.07(m ,1H),2.73-2.85(m,1H),2.39-2.52(m,1H),2.25(s,1H),2.08-2.18(m,1H),1.93-1.98(m,2H),1.80-1.89 (m,1H),1.58-1.63(m,3H),1.49(s,4H),1.38(m,6H). SFC (column: Chiralpak AD-3, 50×4.6mm ID, 3μm; mobile phase: A (supercritical CO 2 ) and B (isopropanol, containing 0.1% isopropylamine); gradient: B%=50～50 %, 3min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=1.367min. ee value: 99%
实施例33
Example 33
步骤1 step 1
在反应瓶中加入化合物13f(50mg,94.87μmol)，化合物1h-2(21.46mg,94.87μmol)，乙酸钾(27.93mg,284.61μmol)，BrettPhos Pd G3(17.20mg,18.97μmol)，二氧六环(2mL)氮气置换三次，100℃搅拌2小时。反应液冷却至室温后加入水(5mL)后用乙酸乙酯萃取(10mL×3)，收集有机相，有机相用饱和食盐水洗涤(4mL×2)，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品用柱层析纯化(二氯甲烷：甲醇＝30:1-10:1)，得到化合物33a。MS m/z:583.4[M+1]+。Add compound 13f (50 mg, 94.87 μmol), compound 1h-2 (21.46 mg, 94.87 μmol), potassium acetate (27.93 mg, 284.61 μmol), BrettPhos Pd G3 (17.20 mg, 18.97 μmol), and dioxane into the reaction bottle. The ring (2 mL) was replaced with nitrogen three times and stirred at 100°C for 2 hours. After the reaction solution was cooled to room temperature, water (5 mL) was added and extracted with ethyl acetate (10 mL × 3). The organic phase was collected, washed with saturated brine (4 mL × 2), dried over anhydrous sodium sulfate, filtered, and reduced pressure. Concentrate to obtain crude product. The crude product was purified by column chromatography (dichloromethane:methanol=30:1-10:1) to obtain compound 33a. MS m/z:583.4[M+1] + .
步骤2Step 2
在反应瓶中加入化合物33a(20mg,34.32μmol)，甲醇(2mL)，多聚甲醛(1.03mg,34.32μmol)，醋酸硼氢化钠(20.16mg,205.94μmol)，冰醋酸(6.18mg,102.97μmol,5.89μL)，25℃搅拌0.5小时。反应液中加入水(4mL)，使用乙酸乙酯萃取(5mL×3)，收集有机相，有机相使用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过薄层硅胶色谱(二氯甲烷：甲醇＝10:1)纯化得到化合物33。MS m/z:597.5[M+1]+。1H NMR(400MHz,CDCl3)δ＝ppm 9.00(s,1H),8.47(s,1H),8.10-8.14(m,1H),8.07(s,1H),7.46(s,1H),6.22(d,J＝5.50Hz,1H),5.41(s,2H),5.10(s,2H),4.53-4.69(m,1H),4.37-4.50(m,1H),3.73(m,1H),3.49-3.63(m,2H),3.33-3.46(m,3H),3.16(q,J＝7.00Hz,4H),2.45(s,3H),1.87-2.05(m,3H),1.54-1.49(m,3H),1.39(d,J＝6.63Hz,6H)。Add compound 33a (20 mg, 34.32 μmol), methanol (2 mL), paraformaldehyde (1.03 mg, 34.32 μmol), sodium acetate borohydride (20.16 mg, 205.94 μmol), and glacial acetic acid (6.18 mg, 102.97 μmol) into the reaction bottle. ,5.89μL), stir at 25°C for 0.5 hours. Water (4 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (5 mL × 3). The organic phase was collected, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by thin-layer silica gel chromatography (dichloromethane:methanol=10:1) to obtain compound 33. MS m/z:597.5[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = ppm 9.00 (s, 1H), 8.47 (s, 1H), 8.10-8.14 (m, 1H), 8.07 (s, 1H), 7.46 (s, 1H), 6.22 (d,J＝5.50Hz,1H),5.41(s,2H),5.10(s,2H),4.53-4.69(m,1H),4.37-4.50(m,1H),3.73(m,1H), 3.49-3.63(m,2H),3.33-3.46(m,3H),3.16(q,J＝7.00Hz,4H),2.45(s,3H),1.87-2.05(m,3H),1.54-1.49( m, 3H), 1.39 (d, J = 6.63Hz, 6H).
实施例34
Example 34
步骤1step 1
将1,4-二氧六环(3mL)，化合物28c(15mg,37.89μmol)，化合物1h-2(6.86mg,30.31μmol)加入长管中，开始搅拌；氮气置换后，依次将乙酸钾(11.16mg,113.66μmol)，BrettPhos Pd G3(6.41mg,7.58μmol)加入其中，升温至90℃，反应4小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相有10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层硅胶色谱分离(乙酸乙酯:甲醇＝30:1)，得到化合物34。MS m/z:586.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.82(s,1H),8.10(d,J＝5.6,1H),7.83(s,1H),7.57(s,1H),6.31(d,J＝5.2,1H),4.91-4.87(m,1H),4.72-4.58(m,2H),4.51-4.47(m,1H),4.31(s,2H),4.08-4.06(m,1H),3.73-3.65(m,1H),3.50-3.28(m,5H),3.04-2.93(m,4H),1.94-1.85(m,2H),1.61(d,J＝6.4,3H),1.56(d,J＝13.6,6H),1.49(d,J＝22.0,3H)。Add 1,4-dioxane (3mL), compound 28c (15mg, 37.89μmol), and compound 1h-2 (6.86mg, 30.31μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate ( 11.16 mg, 113.66 μmol), BrettPhos Pd G3 (6.41 mg, 7.58 μmol) was added, the temperature was raised to 90°C, and the reaction was carried out for 4 hours. After adding 10 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure to obtain crude product. The crude product was separated by thin-layer silica gel chromatography (ethyl acetate:methanol=30:1) to obtain compound 34. MS m/z:586.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.82 (s, 1H), 8.10 (d, J = 5.6, 1H), 7.83 (s, 1H), 7.57 (s, 1H), 6.31 (d, J = 5.2 ,1H),4.91-4.87(m,1H),4.72-4.58(m,2H),4.51-4.47(m,1H),4.31(s,2H),4.08-4.06(m,1H),3.73-3.65 (m,1H),3.50-3.28(m,5H),3.04-2.93(m,4H),1.94-1.85(m,2H),1.61(d,J＝6.4,3H),1.56(d,J＝ 13.6,6H),1.49(d,J=22.0,3H).
实施例35
Example 35
步骤1step 1
将异丙醇(24mL)，化合物29b(1.1g,1.61mmol,50％纯度)，化合物1c(327.80mg,1.77mmol,盐酸盐)(盐酸盐)加入长管中，开始搅拌；然后将二异丙基乙胺(726.01mg,5.62mmol,978.45μL)加入其中，升温至100℃，反应0.5小时。向反应体系中加入40mL水后，加入乙酸乙酯萃取(20mL×2)；得到的有机相用20mL饱和食盐水溶液洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～20:80)，得到化合物35a。MS m/z:342.0[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.85(s,1H),7.33(s,1H),6.12(s,1H),4.77-4.73(m,2H),4.38-4.35(m,2H),3.94(s,3H),3.53-3.48(m,3H),2.99(s,3H)。Add isopropanol (24mL), compound 29b (1.1g, 1.61mmol, 50% purity), compound 1c (327.80mg, 1.77mmol, hydrochloride) (hydrochloride) into a long tube and start stirring; then Diisopropylethylamine (726.01 mg, 5.62 mmol, 978.45 μL) was added, the temperature was raised to 100°C, and the reaction was carried out for 0.5 hours. After adding 40 mL of water to the reaction system, add ethyl acetate and extract (20 mL Get crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~20:80) to obtain compound 35a. MS m/z:342.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.85 (s, 1H), 7.33 (s, 1H), 6.12 (s, 1H), 4.77-4.73 (m, 2H), 4.38-4.35 (m, 2H), 3.94(s,3H),3.53-3.48(m,3H),2.99(s,3H).
步骤2Step 2
将二氯甲烷(20mL)，化合物35a(0.3g,877.67μmol)加入50mL单口瓶中，开始搅拌；然后将温度降至0～5℃，将N-碘代丁二酰亚胺(197.46mg,877.67μmol)溶于0.5mL无水二氯甲烷中，滴加到上述反应液中，反应0.5小时。向反应体系中加入10mL饱和氯化铵溶液后，分液；得到的有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～10:90)，得到化合物35b。MS m/z:467.8[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.77(s,1H),7.47(s,1H),4.72-4.65(m,2H),4.37-4.31(m,2H),3.95(s,3H),3.63(d,J＝7.2,2H),3.38-3.34(m,1H),3.02(s,3H)。Add dichloromethane (20 mL) and compound 35a (0.3 g, 877.67 μmol) into a 50 mL single-neck bottle and start stirring; then lower the temperature to 0~5°C, and add N-iodosuccinimide (197.46 mg, 877.67 μmol) was dissolved in 0.5 mL of anhydrous dichloromethane, and added dropwise to the above reaction solution, and the reaction was carried out for 0.5 hours. After adding 10 mL of saturated ammonium chloride solution to the reaction system, the layers were separated; the obtained organic phase was washed with 10 mL of saturated brine, dried by adding anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~10:90) to obtain compound 35b. MS m/z:467.8[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.77 (s, 1H), 7.47 (s, 1H), 4.72-4.65 (m, 2H), 4.37-4.31 (m, 2H), 3.95 (s, 3H), 3.63(d,J=7.2,2H),3.38-3.34(m,1H),3.02(s,3H).
步骤3Step 3
将1,4-二氧六环(6mL)，水(2.4mL)，化合物35b(150mg,320.71μmol)，异丙烯基硼酸嚬哪醇酯(107.79mg,641.43μmol)加入长管中，开始搅拌；氮气置换后，依次将双(三叔丁基膦)钯(24.59mg,48.11μmol)，碳酸钾(132.98mg,962.14μmol)加入其中，升温至65℃，反应2小时。将反应体系降至室温后，加入30mL饱和氯化铵溶液，然后加入乙酸乙酯萃取(15mL×2)萃取；得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝1:1～1:2.5)，得到化合物35c。MS m/z:382.0[M+1]+。1H NMR(400MHz,DMSO-d6)δ＝8.88(s,1H),7.40(s,1H),5.41(s,1H),4.85(s,1H),4.69-4.65(m,2H),4.35-4.31(m,2H),3.91-3.85(m,3H),3.63-3.61(m,2H),3.39-3.35(m,1H),3.02(s,3H),1.95(s,3H)。Add 1,4-dioxane (6mL), water (2.4mL), compound 35b (150mg, 320.71μmol), isopropenylboronate (107.79mg, 641.43μmol) into the long tube and start stirring. ; After nitrogen replacement, add bis(tri-tert-butylphosphine)palladium (24.59 mg, 48.11 μmol) and potassium carbonate (132.98 mg, 962.14 μmol) in sequence, raise the temperature to 65°C, and react for 2 hours. After the reaction system was brought to room temperature, 30 mL of saturated ammonium chloride solution was added, and then ethyl acetate was added for extraction (15 mL × 2); the obtained organic phase was washed with 15 mL of saturated brine, dried by adding anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure at 45°C to obtain crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:1~1:2.5) to obtain compound 35c. MS m/z:382.0[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δ = 8.88 (s, 1H), 7.40 (s, 1H), 5.41 (s, 1H), 4.85 (s, 1H), 4.69-4.65 (m, 2H), 4.35-4.31(m,2H),3.91-3.85(m,3H),3.63-3.61(m,2H),3.39-3.35(m,1H),3.02(s,3H),1.95(s,3H).
步骤4 Step 4
将甲醇(10mL)，化合物35c(120mg,314.24μmol)加入50mL单口瓶中，开始搅拌；然后将二氧化铂(71.36mg,314.24μmol)加入其中，氢气15psi，室温20℃，反应15小时。反应液直接过滤，滤液在45℃下进行减压浓缩。经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～40:60)，得到化合物35d。MS m/z:384.0[M+1]+。Add methanol (10 mL) and compound 35c (120 mg, 314.24 μmol) into a 50 mL single-neck bottle and start stirring; then add platinum dioxide (71.36 mg, 314.24 μmol), hydrogen 15 psi, room temperature 20°C, and react for 15 hours. The reaction solution was directly filtered, and the filtrate was concentrated under reduced pressure at 45°C. Purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~40:60), compound 35d was obtained. MS m/z:384.0[M+1] + .
步骤5Step 5
将1,4-二氧六环(4mL)，化合物35d(18mg,46.89μmol)，化合物23f(8.49mg,37.51μmol)加入长管中，开始搅拌；氮气置换后，将乙酸钾(13.80mg,140.66μmol)，BrettPhos Pd G3(8.50mg,9.38μmol)加入其中，升温至90℃，反应4小时。向反应体系中加入15mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相加入15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。经薄层色谱分离(乙酸乙酯:甲醇＝30:1)，得到化合物35。MS m/z:574.5[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.81(s,1H),8.40(s,1H),8.08(d,J＝6.0,1H),7.40(s,1H),6.17(d,J＝5.6,1H),4.92-4.78(m,1H),4.71-4.63(m,3H),4.42-4.29(m,3H),3.99(s,3H),3.77-3.46(m,10H),2.99(s,3H),2.07-2.03(m,1H),1.90-1.86(m,1H),1.37(d,J＝6.0Hz,6H)。Add 1,4-dioxane (4mL), compound 35d (18mg, 46.89μmol), and compound 23f (8.49mg, 37.51μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate (13.80mg, 140.66 μmol), BrettPhos Pd G3 (8.50 mg, 9.38 μmol) was added, the temperature was raised to 90°C, and the reaction was carried out for 4 hours. After adding 15 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. After thin layer chromatography separation (ethyl acetate:methanol=30:1), compound 35 was obtained. MS m/z:574.5[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.81 (s, 1H), 8.40 (s, 1H), 8.08 (d, J = 6.0, 1H), 7.40 (s, 1H), 6.17 (d, J = 5.6 ,1H),4.92-4.78(m,1H),4.71-4.63(m,3H),4.42-4.29(m,3H),3.99(s,3H),3.77-3.46(m,10H),2.99(s ,3H),2.07-2.03(m,1H),1.90-1.86(m,1H),1.37(d,J＝6.0Hz,6H).
实施例36
Example 36
步骤1step 1
将二氯甲烷(20mL)，化合物36a(3g,17.42mmol)加入100mL三口瓶中，开始搅拌；然后将三乙胺(4.41g,43.55mmol,6.06mL)加入其中后，将温度降至0～10℃后，将36b(4.01g,22.64mmol,2.75mL)滴加到其中，升至室温20℃，反应0.5小时。向反应体系中加入20mL饱和氯化铵溶液淬灭后，分液；有机相依次用饱和氯化铵溶液(20mL)，饱和食盐水(20mL)洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩，得到化合物36c。1H NMR(400MHz,CDCl3)δ＝5.49(d,J＝7.6Hz,1H),4.28-4.20(m,3H),3.87-3.85(m,2H),3.71-3.63(m,2H),3.19-3.16(m,2H),2.32-2.24(m,2H),1.45-1.44(m,9H)。Add dichloromethane (20mL) and compound 36a (3g, 17.42mmol) into a 100mL three-necked flask and start stirring; then add triethylamine (4.41g, 43.55mmol, 6.06mL) and lower the temperature to 0~ After 10°C, 36b (4.01g, 22.64mmol, 2.75mL) was added dropwise to it, raised to room temperature 20°C, and reacted for 0.5 hours. Add 20 mL of saturated ammonium chloride solution to the reaction system to quench, and then separate the liquids; wash the organic phase with saturated ammonium chloride solution (20 mL) and saturated brine (20 mL) in sequence, add anhydrous sodium sulfate, dry, filter, and remove the filtrate. Concentrate under reduced pressure at 45°C to obtain compound 36c. 1 H NMR (400MHz, CDCl 3 ) δ = 5.49 (d, J = 7.6Hz, 1H), 4.28-4.20 (m, 3H), 3.87-3.85 (m, 2H), 3.71-3.63 (m, 2H), 3.19-3.16(m,2H),2.32-2.24(m,2H),1.45-1.44(m,9H).
步骤2 Step 2
将N,N-二甲基甲酰胺(20mL)，化合物36c(2g,6.39mmol)加入拇指瓶中，开始搅拌；然后将1.8-二氮杂二环[5.4.0]十一烷-7-烯(2.92g,19.18mmol,2.89mL)加入其中，室温20℃，反应2小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相有10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～40:60)，得到化合物36d。MS m/z:220.8[M-56]-。1H NMR(400MHz,CDCl3)δ＝4.24-4.19(m,1H),4.17-4.12(m,2H),4.09-4.05(m,2H),3.37-3.34(m,2H),3.19(d,J＝7.6Hz,2H),2.45-2.38(m,2H),1.44(m,9H)。Add N,N-dimethylformamide (20mL) and compound 36c (2g, 6.39mmol) into the thumb bottle and start stirring; then add 1.8-diazabicyclo[5.4.0]undecane-7- Alkene (2.92g, 19.18mmol, 2.89mL) was added to it, and the reaction was carried out at room temperature 20°C for 2 hours. After adding 10 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. Purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~40:60), compound 36d was obtained. MS m/z:220.8[M-56] - . 1 H NMR (400MHz, CDCl 3 ) δ = 4.24-4.19 (m, 1H), 4.17-4.12 (m, 2H), 4.09-4.05 (m, 2H), 3.37-3.34 (m, 2H), 3.19 (d ,J＝7.6Hz,2H),2.45-2.38(m,2H),1.44(m,9H).
步骤3Step 3
将乙酸乙酯(3mL)，化合物36d(0.3g,1.09mmol)加入50mL单口瓶中，开始搅拌；然后将氯化氢/乙酸乙酯(4M,15mL)缓慢倒入其中，室温20℃，反应1小时。反应液直接过滤，滤饼在40℃下进行减压干燥。得到化合物36e的盐酸盐。MS m/z:176.9[M+1]+。1H NMR(400MHz,DMSO-d6)δ＝4.43-4.33(m,4H),4.27-4.23(m,2H),3.41-3.38(m,2H),3.28-3.25(m,2H),2.44-2.38(m,2H)。Add ethyl acetate (3 mL) and compound 36d (0.3 g, 1.09 mmol) into a 50 mL single-neck bottle and start stirring; then slowly pour hydrogen chloride/ethyl acetate (4 M, 15 mL) into it, and react at room temperature 20°C for 1 hour. . The reaction solution was filtered directly, and the filter cake was dried under reduced pressure at 40°C. The hydrochloride salt of compound 36e was obtained. MS m/z:176.9[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δ = 4.43-4.33 (m, 4H), 4.27-4.23 (m, 2H), 3.41-3.38 (m, 2H), 3.28-3.25 (m, 2H), 2.44 -2.38(m,2H).
步骤4Step 4
将1,4-二氧六环(7.5mL)，化合物15g(100mg,351.40μmol)，碳酸铯(400.72mg,1.23mmol)加入长管中，开始搅拌；氮气置换后，依次将化合物36e(127.06mg,597.37μmol,盐酸盐)，甲磺酸(4,5-双二苯基膦-9,9-二甲基氧杂蒽)(2'-甲胺基-1,1'-联苯-2-基)钯(II)(33.82mg,35.14μmol)加入其中，升温至100℃，反应6小时。向反应体系中加入30mL饱和氯化铵溶液后，加入乙酸乙酯萃取(15mL×2)；有机相有20mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝5:1～1:1)得到化合物36f。MS m/z:380.0[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.11(s,1H),7.85(s,1H),7.48(d,J＝8.0Hz,1H),6.56(d,J＝8.0Hz,1H),4.49-4.41(m,3H),4.27-4.24(m,2H),3.51-3.43(m,3H),3.26-3.22(m,2H),2.48-2.41(m,2H),1.34(dd,J＝6.8Hz,6H)。Add 1,4-dioxane (7.5mL), compound 15g (100mg, 351.40μmol), and cesium carbonate (400.72mg, 1.23mmol) into a long tube and start stirring; after nitrogen replacement, add compound 36e (127.06 mg, 597.37 μmol, hydrochloride), methanesulfonic acid (4,5-bisdiphenylphosphine-9,9-dimethylxanthene) (2'-methylamino-1,1'-biphenyl -2-yl)palladium (II) (33.82 mg, 35.14 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 6 hours. After adding 30mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (15mL×2); wash the organic phase with 20mL saturated brine, add anhydrous sodium sulfate, dry, filter, and reduce the filtrate at 45°C Concentrate under pressure to obtain crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1~1:1) to obtain compound 36f. MS m/z:380.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.11 (s, 1H), 7.85 (s, 1H), 7.48 (d, J = 8.0Hz, 1H), 6.56 (d, J = 8.0Hz, 1H), 4.49 -4.41(m,3H),4.27-4.24(m,2H),3.51-3.43(m,3H),3.26-3.22(m,2H),2.48-2.41(m,2H),1.34(dd,J＝ 6.8Hz, 6H).
步骤5Step 5
将1,4二氧六环(5mL)，化合物36f(30mg,78.97μmol)，化合物23f(16.08mg,71.07μmol)，加入长管中，开始搅拌；氮气置换后，依次将乙酸钾(23.25mg,236.90μmol)，BrettPhos Pd G3(10.74mg,11.85μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入20mL饱和氯化铵溶液后，加入乙酸乙酯萃取(15mL×2)；得到的有机相加入20mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。经薄层色谱法分离(乙酸乙酯:甲醇＝40:1)，得到粗品。向粗品中加入1mL乙腈搅拌15min后，过滤得滤饼为化合物36。MS m/z:570.5[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.06(s,1H),8.57(s,1H),8.08(d,J＝5.6Hz,1H),7.43-7.41(m,2H),6.46(d,J＝8.0Hz,1H),6.22(d,J＝5.6Hz,1H),4.91-4.78(m,1H),4.64-4.57(m,1H),4.50-4.41(m,3H),4.34-4.31(m,1H),4.27-4.23(m,2H),3.86-3.77(m,1H),3.67-3.55(m,3H),3.52(s,3H),3.48-3.45(m,2H),3.24(d,J＝7.6Hz,2H),2.48-2.41(m,2H),2.09-2.01(m,1H),1.91-1.84(m,1H),1.38-1.36(m,6H)。Add 1,4 dioxane (5mL), compound 36f (30mg, 78.97μmol), and compound 23f (16.08mg, 71.07μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate (23.25mg) , 236.90 μmol), BrettPhos Pd G3 (10.74 mg, 11.85 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 3 hours. After adding 20 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (15 mL Concentrate under reduced pressure. Separate by thin layer chromatography (ethyl acetate:methanol=40:1) to obtain crude product. 1 mL of acetonitrile was added to the crude product and stirred for 15 min, then filtered to obtain a filter cake as compound 36. MS m/z:570.5[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.06 (s, 1H), 8.57 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.43-7.41 (m, 2H), 6.46 (d, J＝8.0Hz,1H),6.22(d,J＝5.6Hz,1H),4.91-4.78(m,1H),4.64-4.57(m,1H),4.50-4.41(m,3H),4.34-4.31 (m,1H),4.27-4.23(m,2H),3.86-3.77(m,1H),3.67-3.55(m,3H),3.52(s,3H),3.48-3.45(m,2H),3.24 (d,J＝7.6Hz,2H),2.48-2.41(m,2H),2.09-2.01(m,1H),1.91-1.84(m,1H),1.38-1.36(m,6H).
实施例37
Example 37
步骤1step 1
将1,4二氧六环(5mL)，化合物6d的盐酸盐和7d的盐酸盐混合物(110.53mg)，化合物15g(50mg,175.70μmol)加入长管中，开始搅拌；氮气置换后，将碳酸铯(286.23mg,878.49μmol)，甲磺酸(4,5-双二苯基膦-9,9-二甲基氧杂蒽)(2'-甲胺基-1,1'-联苯-2-基)钯(II)(16.91mg,17.57μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～40:60)，得到化合物37a。MS m/z:377.0[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.11(s,1H),7.87(s,1H),7.50(d,J＝8.0Hz,1H),6.62(d,J＝8.0Hz,1H),5.12-5.11(m,2H),4.90-4.86(m,2H),3.52-3.45(m,1H),3.14-3.09(m,2H),2.71-2.66(m,2H),2.33-2.26(m,2H),1.35(d,J＝6.8Hz,6H)。Add 1,4 dioxane (5mL), the hydrochloride salt of compound 6d and the hydrochloride salt mixture of compound 7d (110.53mg), and compound 15g (50mg, 175.70μmol) into a long tube, and start stirring; after nitrogen replacement, Cesium carbonate (286.23 mg, 878.49 μmol), methanesulfonic acid (4,5-bisdiphenylphosphine-9,9-dimethylxanthene) (2'-methylamino-1,1'-bis Phen-2-yl)palladium (II) (16.91 mg, 17.57 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 3 hours. After adding 10 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~40:60) to obtain compound 37a. MS m/z:377.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.11 (s, 1H), 7.87 (s, 1H), 7.50 (d, J = 8.0Hz, 1H), 6.62 (d, J = 8.0Hz, 1H), 5.12 -5.11(m,2H),4.90-4.86(m,2H),3.52-3.45(m,1H),3.14-3.09(m,2H),2.71-2.66(m,2H),2.33-2.26(m, 2H), 1.35 (d, J = 6.8Hz, 6H).
步骤2Step 2
将1,4-二氧六环(3mL)，化合物37a(15mg,39.80μmol)，化合物23f(8.10mg,35.82μmol)加入长管中，开始搅拌；氮气置换后，将乙酸钾(11.72mg,119.40μmol)，BrettPhos Pd G3(7.22mg,7.96μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入10mL饱和氯化铵溶液以及10mL乙酸乙酯后，分液。得到的水相用乙酸乙酯10mL萃取后，合并有机相，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层色谱法分离(乙酸乙酯:甲醇＝40:1)。得到化合物37。MS m/z:567.2[M+1]+。1H NMR(400MHz,CDCl3)δ＝9.05(s,1H),8.61-8.48(m,1H),8.09(d,J＝6.0Hz,1H),7.46(d,J＝8.0Hz,1H),6.52(d,J＝8.0Hz,1H),6.36-6.25(m,1H),5.15-5.14(m,2H),4.92-4.79(m,3H),4.65-4.55(m,1H),4.37-4.31(m,1H),3.89-3.79(m,1H),3.67-3.54(m,3H),3.52(s,3H),3.11(t,J＝7.2Hz,2H),2.72-2.67(m,2H),2.33-2.26(m,2H),2.09-2.02(m,1H),1.92-1.86(m,1H),1.39-1.36(m,6H)。Add 1,4-dioxane (3mL), compound 37a (15mg, 39.80μmol), and compound 23f (8.10mg, 35.82μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate (11.72mg, 119.40 μmol), BrettPhos Pd G3 (7.22 mg, 7.96 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 3 hours. After adding 10 mL of saturated ammonium chloride solution and 10 mL of ethyl acetate to the reaction system, the liquids were separated. The obtained aqueous phase was extracted with 10 mL of ethyl acetate, the organic phases were combined, dried by adding anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was separated by thin layer chromatography (ethyl acetate:methanol=40:1). Compound 37 was obtained. MS m/z:567.2[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 9.05 (s, 1H), 8.61-8.48 (m, 1H), 8.09 (d, J = 6.0Hz, 1H), 7.46 (d, J = 8.0Hz, 1H) ,6.52(d,J＝8.0Hz,1H),6.36-6.25(m,1H),5.15-5.14(m,2H),4.92-4.79(m,3H),4.65-4.55(m,1H),4.37 -4.31(m,1H),3.89-3.79(m,1H),3.67-3.54(m,3H),3.52(s,3H),3.11(t,J＝7.2Hz,2H),2.72-2.67(m ,2H),2.33-2.26(m,2H),2.09-2.02(m,1H),1.92-1.86(m,1H),1.39-1.36(m,6H).
实施例38、39
Examples 38 and 39
步骤1step 1
在反应瓶中加入化合物15g(200mg,702.79μmol)，化合物13d(264.85mg,702.79μmol,盐酸盐)，二氧六环(6mL)，氮气置换三次，加入碳酸铯(915.94mg,2.81mmol)，甲烷磺酸[9,9-二甲基-4,5-双(二苯基)呫吨](2-甲基氨基-1,1-联苯-2-基)钯(II)(135.27mg,140.56μmol)，100℃搅拌4小时。反应完成后，反应液中加入水(20mL),使用乙酸乙酯萃取(20mL×3)，收集有机相，有机相使用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，滤液减压浓缩得到粗品，粗品通过柱层析纯化(石油醚：乙酸乙酯＝10:0～1:1)得到化合物38a。1H NMR(400MHz,CDCl3)δppm 8.95-9.17(m,1H),7.85(s,1H),7.47(d,J＝7.88Hz,1H),7.37(m,5H),6.42-6.65(m,1H),5.17(s,2H),4.71-4.83(m,1H),4.09-4.25(m,4H),4.03(m,2H),3.91(m,2H),3.43-3.55(m,2H),3.25-3.35(m,1H),3.02-3.16(m,1H),1.35(d,J＝6.75Hz,6H)。Add compound 15g (200mg, 702.79μmol), compound 13d (264.85mg, 702.79μmol, hydrochloride), dioxane (6mL) into the reaction bottle, replace with nitrogen three times, and add cesium carbonate (915.94mg, 2.81mmol) , methanesulfonic acid [9,9-dimethyl-4,5-bis(diphenyl)xanthene](2-methylamino-1,1-biphenyl-2-yl)palladium(II)(135.27 mg, 140.56 μmol), stirred at 100°C for 4 hours. After the reaction is completed, add water (20 mL) to the reaction solution, extract with ethyl acetate (20 mL × 3), collect the organic phase, wash the organic phase with saturated brine (10 mL), dry over anhydrous sodium sulfate, filter, and decompress the filtrate. Concentration gave a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 10:0-1:1) to obtain compound 38a. 1 H NMR (400MHz, CDCl 3 ) δppm 8.95-9.17(m,1H),7.85(s,1H),7.47(d,J＝7.88Hz,1H),7.37(m,5H),6.42-6.65(m ,1H),5.17(s,2H),4.71-4.83(m,1H),4.09-4.25(m,4H),4.03(m,2H),3.91(m,2H),3.43-3.55(m,2H ),3.25-3.35(m,1H),3.02-3.16(m,1H),1.35(d,J＝6.75Hz,6H).
步骤2Step 2
在反应瓶中加入化合物38a(200mg,367.61μmol)，二氯甲烷(10mL)，氮气置换三次加入二乙氨基三氟化硫(65.18mg,404.37μmol,53.43μL)，-30℃搅拌4小时。反应完成后，反应液中加入水(5mL)淬灭，使用二氯甲烷萃取(10mL×3)，收集有机相，有机相使用无水硫酸钠干燥，过滤，滤液减压浓缩得到粗品，粗品通过制备薄层层析(石油醚：乙酸乙酯＝3:1)纯化得到化合物38b。MS m/z:526.2[M+1]+。Add compound 38a (200 mg, 367.61 μmol) and dichloromethane (10 mL) to the reaction bottle, replace with nitrogen three times and add diethylaminosulfur trifluoride (65.18 mg, 404.37 μmol, 53.43 μL), and stir at -30°C for 4 hours. After the reaction is completed, add water (5mL) to the reaction solution to quench it, use dichloromethane to extract (10mL×3), collect the organic phase, dry the organic phase using anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain a crude product, which is passed Compound 38b was purified by preparative thin layer chromatography (petroleum ether: ethyl acetate = 3:1). MS m/z:526.2[M+1] + .
步骤3Step 3
在反应瓶中加入化合物38b(100mg,190.10μmol)，化合物23f(43.01mg,190.10μmol)，二氧六环(5mL)，氮气置换三次加入碳酸铯(185.81mg,570.29μmol)，BrettPhos Pd G3(34.46mg,38.02μmol)，100℃搅拌4小时。反应完成后，反应液中加入水(20mL),使用乙酸乙酯萃取(20mL×3),收集有机相，有机相使用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析(石油醚：乙酸乙酯＝10:0～1:1)纯化得到化合物38c。MS m/z:716.1[M+1]+。Add compound 38b (100 mg, 190.10 μmol), compound 23f (43.01 mg, 190.10 μmol), dioxane (5 mL) to the reaction bottle, replace with nitrogen three times and add cesium carbonate (185.81 mg, 570.29 μmol), BrettPhos Pd G3 ( 34.46 mg, 38.02 μmol), stir at 100°C for 4 hours. After the reaction is completed, add water (20 mL) to the reaction solution, extract with ethyl acetate (20 mL × 3), collect the organic phase, wash the organic phase with saturated brine (10 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Get crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:0-1:1) to obtain compound 38c. MS m/z:716.1[M+1] + .
步骤4 Step 4
在反应瓶中加入化合物38c(50mg,69.85μmol)，二氯甲烷(5mL)，氯化钯(12.39mg,69.85μmol)，三乙基硅烷(16.24mg,139.70μmol)，三乙胺(7.07mg,69.85μmol)，氮气置换三次，25℃搅拌1小时。反应液中加入水(20mL),使用乙酸乙酯萃取(20mL×3),收集有机相，有机相使用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品通过柱层析纯化(二氯甲烷：甲醇＝10:1＝10:0～10:1)纯化得到化合物38d与39d的混合物。MS m/z:582.2,584.2[M+1]+。Add compound 38c (50 mg, 69.85 μmol), dichloromethane (5 mL), palladium chloride (12.39 mg, 69.85 μmol), triethylsilane (16.24 mg, 139.70 μmol), and triethylamine (7.07 mg) into the reaction bottle. , 69.85 μmol), replaced with nitrogen three times, and stirred at 25°C for 1 hour. Water (20 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL × 3), and the organic phase was collected. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:1=10:0~10:1) to obtain a mixture of compounds 38d and 39d. MS m/z:582.2,584.2[M+1] + .
步骤5Step 5
在反应瓶中加入化合物38d与39d的混合物(20mg,34.26μmol)，二氯甲烷(4mL)，醋酸硼氢化钠(43.57mg,205.58μmol),乙醛(3.77mg,34.26μmol,40％纯度),冰醋酸(28.38mg,102.79μmol)，氮气置换三次，25℃搅拌1小时。反应完成后，反应液中加入水(5mL)使用二氯甲烷萃取(5mL×3)，收集有机相，有机相使用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品经制备高效液相色谱分离(色谱柱:Waters Xbridge Prep OBD C18 150×40mm×10μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:40-70％)得到化合物38和化合物39。Add the mixture of compounds 38d and 39d (20 mg, 34.26 μmol), dichloromethane (4 mL), sodium acetate borohydride (43.57 mg, 205.58 μmol), acetaldehyde (3.77 mg, 34.26 μmol, 40% purity) into the reaction bottle. , glacial acetic acid (28.38 mg, 102.79 μmol), replaced with nitrogen three times, and stirred at 25°C for 1 hour. After the reaction is completed, add water (5 mL) to the reaction solution and extract with dichloromethane (5 mL × 3). Collect the organic phase. Wash the organic phase with saturated brine (5 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain Crude. The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Waters 70%) to obtain compound 38 and compound 39.
化合物38：MS m/z:610.6[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.05(s,1H),8.45-8.68(m,1H),8.10(d,J＝6.13Hz,1H),7.47(d,J＝7.88Hz,1H),6.53(d,J＝7.88Hz,1H),6.22-6.40(m,1H),5.18(s,2H),4.80-4.95(m,3H),4.55-4.72(m,1H),4.31-4.44(m,1H),3.77-3.95(m,1H),3.55-3.72(m,3H),3.45-3.54(m,6H),3.22(s,4H),2.66(q,J＝7.17Hz,2H),2.03-2.16(m,1H),1.86-1.95(m,1H),1.37(dd,J＝6.69,3.06Hz,6H),1.16(t,J＝7.07Hz,3H)。Compound 38: MS m/z: 610.6[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.05 (s, 1H), 8.45-8.68 (m, 1H), 8.10 (d, J = 6.13Hz, 1H), 7.47 (d, J = 7.88Hz, 1H), 6.53(d,J＝7.88Hz,1H),6.22-6.40(m,1H),5.18(s,2H),4.80-4.95(m,3H),4.55-4.72(m,1H),4.31-4.44( m,1H),3.77-3.95(m,1H),3.55-3.72(m,3H),3.45-3.54(m,6H),3.22(s,4H),2.66(q,J＝7.17Hz,2H) ,2.03-2.16(m,1H),1.86-1.95(m,1H),1.37(dd,J＝6.69,3.06Hz,6H),1.16(t,J＝7.07Hz,3H).
化合物39：MS m/z:612.6[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.50-8.62(m,1H),8.09(d,J＝5.88Hz,1H),7.61-7.81(m,1H),7.44(d,J＝8.00Hz,1H),6.45(d,J＝8.13Hz,1H),6.22-6.36(m,1H),4.76-4.96(m,1H),4.61(m,1H),4.38-4.49(m,2H),4.30-4.38(m,1H),4.26(dd,J＝7.75,6.13Hz,1H),3.95(t,J＝6.50Hz,1H),3.77-3.90(m,1H),3.54-3.68(m,3H),3.52(s,3H),3.34-3.47(m,2H),3.03-3.20(m,4H),2.92(m,1H),2.70(m,1H),2.61(m,2H),2.06(m,1H),1.86-1.92(m,1H),1.37(dd,J＝6.75,2.75Hz,6H),1.11(t,J＝7.13Hz,3H)。Compound 39: MS m/z: 612.6[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07 (s, 1H), 8.50-8.62 (m, 1H), 8.09 (d, J = 5.88Hz, 1H), 7.61-7.81 (m, 1H), 7.44 (d ,J＝8.00Hz,1H),6.45(d,J＝8.13Hz,1H),6.22-6.36(m,1H),4.76-4.96(m,1H),4.61(m,1H),4.38-4.49( m,2H),4.30-4.38(m,1H),4.26(dd,J＝7.75,6.13Hz,1H),3.95(t,J＝6.50Hz,1H),3.77-3.90(m,1H),3.54 -3.68(m,3H),3.52(s,3H),3.34-3.47(m,2H),3.03-3.20(m,4H),2.92(m,1H),2.70(m,1H),2.61(m ,2H),2.06(m,1H),1.86-1.92(m,1H),1.37(dd,J＝6.75,2.75Hz,6H),1.11(t,J＝7.13Hz,3H).
实施例40
Example 40
步骤1step 1
将异丙醇(12mL)，化合物14e(250mg,653.18μmol)，化合物1c(145.53mg,783.82μmol,盐酸盐)加入长管中，开始搅拌；然后将N,N-二异丙基乙胺(253.25mg,1.96mmol,341.31μL)加入其中，升温至100℃，反应2小时。向反应体系中加入30mL水后，加入乙酸乙酯萃取(20mL×2)；得到的有机相用20mL饱和食盐水溶液洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经 硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～50:50)，得到化合物40a。MS m/z:382.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.84(s,1H),7.32(s,1H),6.17(s,1H),5.07(s,1H),4.97(s,1H),4.76-4.72(m,4H),4.38-4.34(m,2H),3.53-3.48(m,3H),3.00(s,3H),1.84(s,3H)。Add isopropyl alcohol (12 mL), compound 14e (250 mg, 653.18 μmol), and compound 1c (145.53 mg, 783.82 μmol, hydrochloride) into a long tube and start stirring; then add N,N-diisopropylethylamine (253.25 mg, 1.96 mmol, 341.31 μL) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. After adding 30 mL of water to the reaction system, add ethyl acetate for extraction (20 mL × 2); wash the obtained organic phase with 20 mL of saturated saline solution, add anhydrous sodium sulfate, dry, filter, and concentrate the filtrate under reduced pressure at 45°C. Get crude product. Crude Classics Separate and purify by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~50:50) to obtain compound 40a. MS m/z:382.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.84 (s, 1H), 7.32 (s, 1H), 6.17 (s, 1H), 5.07 (s, 1H), 4.97 (s, 1H), 4.76-4.72 ( m,4H),4.38-4.34(m,2H),3.53-3.48(m,3H),3.00(s,3H),1.84(s,3H).
步骤2Step 2
将二氯甲烷(16mL)，化合物40a(230mg,602.29μmol)加入拇指瓶中，开始搅拌；然后将温度降至0～5℃后，将N-碘代丁二酰亚胺(128.73mg,572.17μmol)溶于3mL二氯甲烷中后，缓慢滴加到其中，反应0.5小时。向反应体系中加入15mL水后，分液；得到的有机相用15mL饱和食盐水溶液洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～50:50)，得到化合物40b。MS m/z:507.9[M+1]+。Add dichloromethane (16 mL) and compound 40a (230 mg, 602.29 μmol) into the thumb bottle and start stirring; then lower the temperature to 0~5°C, add N-iodosuccinimide (128.73 mg, 572.17 μmol) was dissolved in 3 mL of methylene chloride, and then slowly added dropwise to it, and the reaction was carried out for 0.5 hours. After adding 15 mL of water to the reaction system, the layers were separated; the obtained organic phase was washed with 15 mL of saturated saline solution, dried by adding anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C. After separation and purification by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~50:50), compound 40b was obtained. MS m/z:507.9[M+1] + .
步骤3Step 3
将1,4-二氧六环(1mL)，化合物40b(50mg,98.47μmol)加入长管中，开始搅拌；氮气置换后，然后将甲酸钠(16.74mg,246.17μmol,13.29μL)，四丁基氯化铵(41.05mg,147.70μmol,41.30μL)，乙酸钾(38.66mg,393.88μmol)，醋酸钯(4.42mg,19.69μmol)加入其中，升温至90℃，反应4小时。向反应体系中加入10mL水以及10mL乙酸乙酯后，分液；得到的有机相加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩。经硅胶柱层析分离纯化(石油醚:乙酸乙酯＝100:0～10:90)，得到化合物40c。MS m/z:382.1[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.87(s,1H),7.39(s,1H),4.76-4.72(m,2H),4.37-4.32(m,4H),3.47-3.45(m,2H),3.01-2.97(m,4H),1.52(s,6H)。Add 1,4-dioxane (1mL) and compound 40b (50mg, 98.47μmol) into the long tube and start stirring; after nitrogen replacement, add sodium formate (16.74mg, 246.17μmol, 13.29μL), tetrabutyl Ammonium chloride (41.05 mg, 147.70 μmol, 41.30 μL), potassium acetate (38.66 mg, 393.88 μmol), and palladium acetate (4.42 mg, 19.69 μmol) were added, and the temperature was raised to 90°C and reacted for 4 hours. After adding 10 mL of water and 10 mL of ethyl acetate to the reaction system, the liquids were separated; the obtained organic phase was dried by adding anhydrous sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure at 45°C. After separation and purification by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~10:90), compound 40c was obtained. MS m/z:382.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.87 (s, 1H), 7.39 (s, 1H), 4.76-4.72 (m, 2H), 4.37-4.32 (m, 4H), 3.47-3.45 (m, 2H) ),3.01-2.97(m,4H),1.52(s,6H).
步骤4Step 4
将1,4-二氧六环(3mL)，化合物23f(9.48mg,41.90μmol)，化合物40c(20mg,52.37μmol)加入长管中，开始搅拌；氮气置换后，将乙酸钾(15.42mg,157.12μmol)，BrettPhos Pd G3(9.50mg,10.47μmol)加入其中，升温至90℃，反应3小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相有10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层色谱(TLC)分离(展开剂为乙酸乙酯:甲醇＝30:1)，得到化合物40。MS m/z:572.4[M+1]+。1H NMR(400MHz,CDCl3)δ＝8.82(s,1H),8.10(d,J＝5.6,1H),7.88(s,1H),7.42(s,1H),6.29(d,J＝5.6,1H),4.89-4.70(m,3H),4.65-4.58(m,1H),4.36-4.29(m,5H),3.77-3.68(m,1H),3.63-3.54(m,2H),3.51(s,3H),3.47-3.42(m,3H),2.98(s,3H),2.05-1.95(m,1H),1.88-1.81(m,1H),1.57(s,6H)。Add 1,4-dioxane (3mL), compound 23f (9.48mg, 41.90μmol), and compound 40c (20mg, 52.37μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate (15.42mg, 157.12 μmol), BrettPhos Pd G3 (9.50 mg, 10.47 μmol) was added, the temperature was raised to 90°C, and the reaction was carried out for 3 hours. After adding 10 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure to obtain crude product. The crude product was separated by thin layer chromatography (TLC) (the developing solvent was ethyl acetate:methanol=30:1) to obtain compound 40. MS m/z:572.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.82 (s, 1H), 8.10 (d, J = 5.6, 1H), 7.88 (s, 1H), 7.42 (s, 1H), 6.29 (d, J = 5.6 ,1H),4.89-4.70(m,3H),4.65-4.58(m,1H),4.36-4.29(m,5H),3.77-3.68(m,1H),3.63-3.54(m,2H),3.51 (s,3H),3.47-3.42(m,3H),2.98(s,3H),2.05-1.95(m,1H),1.88-1.81(m,1H),1.57(s,6H).
实施例41
Example 41
步骤1step 1
在反应瓶中加入化合物36a(1g,5.81mmol)，二氯甲烷(30mL)，三乙胺(1.47g,14.52mmol)氮气置换三次，0-10℃温度滴加化合物41a(1.33g,6.97mmol)，升温至25℃，搅拌1小时。加入水(5mL)淬灭，二氯甲烷萃取(10mL×2)，收集有机相，无水硫酸钠干燥，过滤，减压浓缩得到粗品，快速柱层析纯化(二氯甲烷：甲醇＝10：0～10:1)纯化得到41b。1H NMR(400MHz,CDCl3)δppm 4.90(m,1H),4.16-4.34(m,2H),3.77-3.90(m,2H),3.58(t,J＝5.88Hz,2H),2.99-3.09(m,2H),1.88-2.07(m,4H),1.45(s,9H)。Add compound 36a (1g, 5.81mmol), dichloromethane (30mL), and triethylamine (1.47g, 14.52mmol) to the reaction bottle and replace it with nitrogen three times. Compound 41a (1.33g, 6.97mmol) is added dropwise at 0-10°C. ), raise the temperature to 25°C, and stir for 1 hour. Add water (5mL) to quench, extract with dichloromethane (10mL×2), collect the organic phase, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain crude product, purify by flash column chromatography (dichloromethane:methanol=10: 0～10:1) to obtain 41b. 1 H NMR (400MHz, CDCl 3 ) δppm 4.90 (m, 1H), 4.16-4.34 (m, 2H), 3.77-3.90 (m, 2H), 3.58 (t, J = 5.88Hz, 2H), 2.99-3.09 (m,2H),1.88-2.07(m,4H),1.45(s,9H).
步骤2Step 2
在反应瓶中加入化合物41b(1g,3.06mmol)，N,N-二甲基甲酰胺(10mL)，1.8-二氮杂二环[5.4.0]十一烷-7-烯(1.40g,9.18mmol)，氮气置换三次，25℃搅拌2小时。加入水(5mL)淬灭，乙酸乙酯萃取(10mL×2)，有机相使用饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品，通过快速柱层析纯化(石油醚：乙酸乙酯＝10：0～1：1)得到化合物41c。1H NMR(400MHz,CDCl3)δ＝4.50-4.65(m,1H),4.10(d,J＝7.88Hz,2H),3.98(dd,J＝9.69,5.82Hz,2H),3.35-3.43(m,2H),3.02-3.10(m,2H),2.16-2.28(m,2H),1.70-1.81(m,2H),1.44(s,9H)。Add compound 41b (1g, 3.06mmol), N,N-dimethylformamide (10mL), 1.8-diazabicyclo[5.4.0]undecane-7-ene (1.40g, 9.18 mmol), replaced with nitrogen three times, and stirred at 25°C for 2 hours. Add water (5 mL) to quench, extract with ethyl acetate (10 mL × 2), wash the organic phase with saturated brine (10 mL × 2), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product, which is then subjected to flash column chromatography. Purification (petroleum ether: ethyl acetate = 10:0-1:1) gave compound 41c. 1 H NMR (400MHz, CDCl 3 ) δ = 4.50-4.65 (m, 1H), 4.10 (d, J = 7.88Hz, 2H), 3.98 (dd, J = 9.69, 5.82Hz, 2H), 3.35-3.43 ( m,2H),3.02-3.10(m,2H),2.16-2.28(m,2H),1.70-1.81(m,2H),1.44(s,9H).
步骤3Step 3
在反应瓶中加入化合物41c(0.7g,2.41mmol)，氯化氢/乙酸乙酯(10mL，4M)，25℃搅拌2小时。反应液进行减压浓缩得到化合物41d的盐酸盐。1H NMR(400MHz,CD3OD)δppm 4.66(t,J＝7.94Hz,1H),4.28(m,2H),4.18(m,2H),3.35-3.42(m,2H),3.12-3.21(m,2H),2.16(m,2H),1.61-1.74(m,2H)。Compound 41c (0.7g, 2.41mmol) and hydrogen chloride/ethyl acetate (10mL, 4M) were added to the reaction flask, and stirred at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the hydrochloride salt of compound 41d. 1 H NMR (400MHz, CD 3 OD) δppm 4.66 (t, J＝7.94Hz, 1H), 4.28 (m, 2H), 4.18 (m, 2H), 3.35-3.42 (m, 2H), 3.12-3.21 ( m,2H),2.16(m,2H),1.61-1.74(m,2H).
步骤4Step 4
在反应瓶中加入化合物15g(200mg,702.79μmol)，化合物41d(191.21mg,843.35μmol,盐酸盐)，二氧六环(6mL)，氮气置换三次加入碳酸铯(915.94mg,2.81mmol)，Xantphos-Pd-G4(135.27mg,140.56μmol)，100℃搅拌4小时。加入水(5mL)淬灭，乙酸乙酯萃取(10mL×2)，收集有机相用饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品，通过快速柱层析纯化(石油醚：乙酸乙酯＝10：0～1：1)得到化合物41e。MS m/z:394.4[M+1]+。Add compound 15g (200mg, 702.79μmol), compound 41d (191.21mg, 843.35μmol, hydrochloride), dioxane (6mL) into the reaction bottle, replace with nitrogen three times and add cesium carbonate (915.94mg, 2.81mmol). Xantphos-Pd-G4 (135.27 mg, 140.56 μmol), stirred at 100°C for 4 hours. Add water (5 mL) to quench, extract with ethyl acetate (10 mL × 2), collect the organic phase and wash with saturated brine (10 mL × 2), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product, which is passed through flash column layer. Compound 41e was obtained by analytical purification (petroleum ether: ethyl acetate = 10:0-1:1). MS m/z:394.4[M+1] + .
步骤5Step 5
在反应瓶中加入化合物41e(140mg,355.39μmol)，化合物23f(64.33mg,284.31μmol)，二氧六环(4mL)，氮气置换三次加入碳酸铯(347.38mg,1.07mmol)，BrettPhos Pd G3(64.43mg,71.08μmol)，100℃搅拌2小时。加入水(5mL)淬灭，乙酸乙酯萃取(10mL×2)，收集有机相，用饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品，通过柱层析纯化(二氯甲烷：甲醇＝10:0～10：1)得到化合物41。MS m/z:584.5[M+1]+。1H NMR(400MHz,CDCl3)δppm 9.06(s,1H),8.57(s,1H),8.08(d,J＝5.75Hz,1H),7.46(s,1H),7.42(d,J＝8.00Hz,1H),6.47(d,J＝8.00Hz,1H),6.16-6.29(m,1H),4.71-4.95(m,2H),4.54-4.65(m,1H),4.42(t,J＝7.57Hz,2H),4.31(m,1H),4.11(t,J＝7.00Hz,2H),3.73-3.92(m,1H),3.56-3.70(m,3H),3.52(s,3H),3.44-3.50(m,2H),3.06-3.14(m,2H),2.25(m,2H),2.05(m,1H),1.87(ddd,J＝9.91,6.72,3.38Hz,1H),1.77(m,2H),1.37(dd,J＝6.75,2.25Hz,6H)。Add compound 41e (140 mg, 355.39 μmol), compound 23f (64.33 mg, 284.31 μmol), dioxane (4 mL) to the reaction flask, replace with nitrogen three times and add cesium carbonate (347.38 mg, 1.07 mmol), BrettPhos Pd G3 ( 64.43 mg, 71.08 μmol), stir at 100°C for 2 hours. Add water (5 mL) to quench, extract with ethyl acetate (10 mL × 2), collect the organic phase, wash with saturated brine (10 mL × 2), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product, which is passed through the column layer. Compound 41 was obtained by analytical purification (dichloromethane:methanol=10:0~10:1). MS m/z:584.5[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.06 (s, 1H), 8.57 (s, 1H), 8.08 (d, J = 5.75Hz, 1H), 7.46 (s, 1H), 7.42 (d, J = 8.00 Hz,1H),6.47(d,J＝8.00Hz,1H),6.16-6.29(m,1H),4.71-4.95(m,2H),4.54-4.65(m,1H),4.42(t,J＝ 7.57Hz,2H),4.31(m,1H),4.11(t,J＝7.00Hz,2H),3.73-3.92(m,1H),3.56-3.70(m,3H),3.52(s,3H), 3.44-3.50(m,2H),3.06-3.14(m,2H),2.25(m,2H),2.05(m,1H),1.87(ddd,J＝9.91,6.72,3.38Hz,1H),1.77( m, 2H), 1.37 (dd, J = 6.75, 2.25Hz, 6H).
实施例42
Example 42
步骤1step 1
在反应瓶中加入化合物38d与39d的混合物(40mg,68.53μmol)，二氯甲烷(2mL)，丙酮(7.96mg,137.05μmol)，冰醋酸(12.35mg,205.58μmol)，醋酸硼氢化钠(87.14mg,411.16μmol)，氮气置换三次，25℃搅拌1小时。反应完成后，反应液中加入水(5mL)使用二氯甲烷萃取(5mL×3)，收集有机相，有机相使用饱和食盐水(5mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品经制备高效液相色谱分离(色谱柱:Waters Xbridge Prep OBD C18 150×40mm×10μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:45-65％)得到化合物42。MS ESI m/z:626.6[M+H]+。1H NMR(400MHz,CDCl3)δppm9.08(s,1H),8.55(s,1H),8.08(d,J＝5.75Hz,1H),7.53-7.64(m,1H),7.43(d,J＝8.00Hz,1H),6.44(d,J＝8.00Hz,1H),6.19-6.32(m,1H),4.74-4.95(m,1H),4.61(dt,J＝13.23,6.71Hz,1H),4.44(dt,J＝12.88,7.63Hz,2H),4.29-4.38(m,1H),4.25(t,J＝7.19Hz,1H),3.93(t,J＝6.88Hz,1H),3.75-3.88(m,1H),3.55-3.70(m,3H),3.52(s,3H),3.39-3.48(m,1H),3.27-3.37(m,1H),3.04-3.16(m,3H),2.97(m,3H),2.72-2.83(m,1H),2.01-2.09(m,1H),1.88(m,1H),1.37(dd,J＝6.75,2.50Hz,6H),1.06(t,J＝6.88Hz,6H)。Add the mixture of compounds 38d and 39d (40 mg, 68.53 μmol), dichloromethane (2 mL), acetone (7.96 mg, 137.05 μmol), glacial acetic acid (12.35 mg, 205.58 μmol), and sodium borohydride acetate (87.14) into the reaction bottle. mg, 411.16 μmol), replaced with nitrogen three times, and stirred at 25°C for 1 hour. After the reaction is completed, add water (5 mL) to the reaction solution and extract with dichloromethane (5 mL × 3). Collect the organic phase. Wash the organic phase with saturated brine (5 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain Crude. The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Waters Xbridge Prep OBD C18 150×40mm×10μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% ammonia bicarbonate); gradient: B%: 45- 65%) to obtain compound 42. MS ESI m/z:626.6[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm9.08 (s, 1H), 8.55 (s, 1H), 8.08 (d, J = 5.75Hz, 1H), 7.53-7.64 (m, 1H), 7.43 (d, J＝8.00Hz,1H),6.44(d,J＝8.00Hz,1H),6.19-6.32(m,1H),4.74-4.95(m,1H),4.61(dt,J＝13.23,6.71Hz,1H ),4.44(dt,J＝12.88,7.63Hz,2H),4.29-4.38(m,1H),4.25(t,J＝7.19Hz,1H),3.93(t,J＝6.88Hz,1H),3.75 -3.88(m,1H),3.55-3.70(m,3H),3.52(s,3H),3.39-3.48(m,1H),3.27-3.37(m,1H),3.04-3.16(m,3H) ,2.97(m,3H),2.72-2.83(m,1H),2.01-2.09(m,1H),1.88(m,1H),1.37(dd,J＝6.75,2.50Hz,6H),1.06(t ,J＝6.88Hz,6H).
实施例43
Example 43
步骤1step 1
将N,N-二甲基甲酰胺(3mL)，化合物1h-2(0.1g,441.99μmol)加入拇指瓶中，开始搅拌；然后将温度降至0～5℃后，将咪唑(60.18mg,883.98μmol)加入其中，反应15min后，将叔丁基二甲基氯硅烷(99.92mg,662.98μmol)滴加到其中，升至室温20℃，反应20小时。向反应体系中加入15mL饱和氯化铵溶液后，加入乙酸乙酯(10mL×2次)萃取；得到的有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～35:65)，得到化合物43a。MS ESI m/z:341.4[M+H]+。Add N,N-dimethylformamide (3mL) and compound 1h-2 (0.1g, 441.99μmol) into the thumb bottle and start stirring; then lower the temperature to 0~5℃, add imidazole (60.18mg, 883.98 μmol) was added to it, and after reacting for 15 minutes, tert-butyldimethylsilyl chloride (99.92 mg, 662.98 μmol) was added dropwise, and the mixture was raised to room temperature 20°C and allowed to react for 20 hours. After adding 15 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate (10 mL Concentrate under reduced pressure to obtain crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0~35:65) to obtain compound 43a. MS ESI m/z:341.4[M+H]+.
步骤2Step 2
将二氯甲烷(10mL)，化合物43a(90mg,264.31μmol)，加入长管中，开始搅拌；然后依次将N,N-二异丙基乙胺(204.96mg,1.59mmol)，2-(三甲基硅)乙氧基甲基氯(264.39mg,1.59mmol)加入其中，升温至40℃，反应20小时。向反应体系中加入20mL饱和氯化铵溶液以及15mL二氯甲烷后分液；得到的有机相加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～30:70)，得到化合物43b。1H NMR(400MHz,DMSO-d6)δppm 7.96(d,J＝5.6Hz,1H),6.08(d,J＝5.6Hz,1H),5.1-4.60(m,4H),4.49-4.35(m,2H),3.82-3.74(m,1H),3.52(t,J＝8.0Hz,4H),3.25-3.16(m,2H),1.66-1.65(m,2H),1.29(d,J＝21.6Hz,3H),0.88-0.83(m,13H),0.15-0.07(m,6H),-0.01--0.04(m,18H)。Add dichloromethane (10 mL) and compound 43a (90 mg, 264.31 μmol) into a long tube and start stirring; then add N,N-diisopropylethylamine (204.96 mg, 1.59 mmol), 2-(tris Methyl silicon) ethoxymethyl chloride (264.39 mg, 1.59 mmol) was added, the temperature was raised to 40°C, and the reaction was carried out for 20 hours. Add 20 mL of saturated ammonium chloride solution and 15 mL of methylene chloride to the reaction system and separate the layers; add anhydrous sodium sulfate to the obtained organic phase, dry it, filter it, and concentrate the filtrate under reduced pressure at 45°C to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0~30:70) to obtain compound 43b. 1H NMR(400MHz,DMSO-d6)δppm 7.96(d,J＝5.6Hz,1H),6.08(d,J＝5.6Hz,1H),5.1-4.60(m,4H),4.49-4.35(m,2H ),3.82-3.74(m,1H),3.52(t,J＝8.0Hz,4H),3.25-3.16(m,2H),1.66-1.65(m,2H),1.29(d,J＝21.6Hz, 3H),0.88-0.83(m,13H),0.15-0.07(m,6H),-0.01--0.04(m,18H).
步骤3Step 3
将四氢呋喃(5mL)，化合物43b(70mg,116.47μmol)加入拇指瓶中，开始搅拌；然后将四丁基氟化铵(1M,582.33μL)滴加到其中，室温20℃，反应2小时。向反应体系中加入20mL饱和氯化铵溶液以及20mL乙酸乙酯后，分液；得到的有机相依次用饱和氯化铵溶液(20mL×2)，饱和食盐水(20mL)洗涤 后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。得到化合物43c。Add tetrahydrofuran (5 mL) and compound 43b (70 mg, 116.47 μmol) into the thumb bottle and start stirring; then add tetrabutylammonium fluoride (1 M, 582.33 μL) dropwise into it, and react at room temperature 20°C for 2 hours. After adding 20 mL saturated ammonium chloride solution and 20 mL ethyl acetate to the reaction system, the liquids were separated; the obtained organic phase was washed with saturated ammonium chloride solution (20 mL × 2) and saturated brine (20 mL). Then, add anhydrous sodium sulfate to dry, filter, and concentrate the filtrate under reduced pressure at 45°C. Compound 43c was obtained.
步骤4Step 4
将N,N-二甲基甲酰胺(9mL)，化合物43c(60mg,123.26μmol)加入拇指瓶中，开始搅拌；然后将温度降至0～5℃后，将氢化钠(5.42mg,135.59μmol,60％纯度)加入其中后，反应15min，然后将碘甲烷(17.50mg,123.26μmol,7.67μL)加入其中，继续反应1小时。向反应体系中加入20mL饱和氯化铵溶液后，加入乙酸乙酯(10mL×2)萃取；得到的有机相用20mL饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在45℃下进行减压浓缩。得到化合物43d。Add N,N-dimethylformamide (9mL) and compound 43c (60mg, 123.26μmol) into the thumb bottle and start stirring; then lower the temperature to 0~5℃, add sodium hydride (5.42mg, 135.59μmol , 60% purity) was added to it, reacted for 15 minutes, then methyl iodide (17.50 mg, 123.26 μmol, 7.67 μL) was added, and the reaction continued for 1 hour. After adding 20 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate (10 mL Concentrate under reduced pressure. Compound 43d was obtained.
步骤5Step 5
将乙醇(5mL)，化合物43d(55mg,109.83μmol)加入拇指瓶中，开始搅拌；然后将盐酸(6M,1.10mL)加入其中，升温至75℃，反应1小时。反应液在45℃下进行减压浓缩。得到化合物43e的盐酸盐。MS ESI m/z:241.3[M+H]+。Add ethanol (5 mL) and compound 43d (55 mg, 109.83 μmol) into the thumb bottle and start stirring; then add hydrochloric acid (6 M, 1.10 mL), raise the temperature to 75°C, and react for 1 hour. The reaction solution was concentrated under reduced pressure at 45°C. The hydrochloride salt of compound 43e was obtained. MS ESI m/z:241.3[M+H]+.
步骤6Step 6
将1,4二氧六环(4mL)，化合物24a(25mg,65.98μmol)，化合物43e的盐酸盐(18.26mg,65.98μmol)加入长管中，开始搅拌；氮气置换后，然后依次将碳酸铯(68.79mg,211.13μmol)，甲磺酸(2-二环己基膦基-3,6-二甲氧基-2,4,6-三异丙基-1,1-联苯)(2-氨基-1,1-联苯-2-基)钯(II)(11.96mg,13.20μmol)加入其中，升温至100℃，反应2小时。向反应体系中加入15mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经薄层色谱分离(展开剂为乙酸乙酯：甲醇＝40:1)，得到化合物43。MS ESI m/z:583.5[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.48(s,1H),8.08(d,J＝6.0Hz,1H),7.53-7.39(m,2H),6.42(d,J＝8.0Hz,1H),6.31-6.23(m,1H),4.69-4.57(m,2H),4.46-4.38(m,2H),4.16(dd,J＝8.0and5.2Hz,1H),3.99-3.96(m,1H),3.60-3.53(m,1H),3.51-3.39(m,6H),3.35-3.26(m,1H),3.24-3.11(m,2H),3.07-3.01(m,1H),2.52-2.44(m,1H),2.32-2.11(m,2H),2.02-1.86(m,3H),1.51(d,J＝21.6,3H),1.37(d,J＝6.8Hz,6H)。Add 1,4 dioxane (4 mL), compound 24a (25 mg, 65.98 μmol), and the hydrochloride of compound 43e (18.26 mg, 65.98 μmol) into a long tube, and start stirring; after nitrogen replacement, add carbonic acid in turn Cesium (68.79 mg, 211.13 μmol), methanesulfonic acid (2-dicyclohexylphosphino-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl) (2 -Amino-1,1-biphenyl-2-yl)palladium (II) (11.96 mg, 13.20 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2 hours. After adding 15 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure to obtain crude product. The crude product was separated by thin layer chromatography (developing solvent: ethyl acetate: methanol = 40:1) to obtain compound 43. MS ESI m/z:583.5[M+H]+. 1H NMR (400MHz, CDCl3) δppm 9.07 (s, 1H), 8.48 (s, 1H), 8.08 (d, J = 6.0Hz, 1H), 7.53-7.39 (m, 2H), 6.42 (d, J = 8.0 Hz,1H),6.31-6.23(m,1H),4.69-4.57(m,2H),4.46-4.38(m,2H),4.16(dd,J＝8.0and5.2Hz,1H),3.99-3.96( m,1H),3.60-3.53(m,1H),3.51-3.39(m,6H),3.35-3.26(m,1H),3.24-3.11(m,2H),3.07-3.01(m,1H), 2.52-2.44(m,1H),2.32-2.11(m,2H),2.02-1.86(m,3H),1.51(d,J=21.6,3H),1.37(d,J=6.8Hz,6H).
实施例44
Example 44
步骤1step 1
在反应瓶中加入化合物44a(20g,153.68mmol,19.05mL)，四氢呋喃(300mL)，碘代异丙烷(26.12g,153.68mmol,15.37mL)，碳酸钾(84.96g,614.72mmol)，氮气置换三次，70℃搅拌12小时。反应液中加入水(100mL)，使用乙酸乙酯萃取(100mL×2)，收集有机相，有机相使用饱和食盐水洗涤(50mL)，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品用柱层析纯化(石油醚：乙酸乙酯＝10:0～3:1)纯化得到化合 物44b。1H NMR(400MHz,CDCl3)δppm 3.68(s,3H),3.22(d,J＝12.00Hz,1H),2.35-2.63(m,3H),1.03(t,J＝8.00Hz,3H),0.93(d,J＝8.00Hz,3H),0.88(d,J＝4.00Hz,3H)。Add compound 44a (20g, 153.68mmol, 19.05mL), tetrahydrofuran (300mL), isopropane iodide (26.12g, 153.68mmol, 15.37mL), potassium carbonate (84.96g, 614.72mmol) into the reaction flask, and replace with nitrogen three times. , stirred at 70°C for 12 hours. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL × 2). The organic phase was collected, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:0~3:1) to obtain the compound. Thing 44b. 1 H NMR (400MHz, CDCl 3 ) δppm 3.68 (s, 3H), 3.22 (d, J = 12.00Hz, 1H), 2.35-2.63 (m, 3H), 1.03 (t, J = 8.00Hz, 3H), 0.93(d,J=8.00Hz,3H), 0.88(d,J=4.00Hz,3H).
步骤2Step 2
在100mL三口反应瓶中加入化合物44b(3.58g,20.81mmol)，四氢呋喃(30mL)氮气置换三次降温至0℃加入钠氢(1.39g,34.68mmol,60％纯度)，0℃搅拌0.5小时加入4，6-二氯烟腈(3g,17.34mmol)，25℃搅拌10小时。反应完成后反应液加入到饱和氯化铵水溶液(40mL)淬灭，使用乙酸乙酯萃取(50mL×2)，收集有机相，有机相使用饱和食盐水(40mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到产物。产物通过柱层析纯化(石油醚：乙酸乙酯＝10:0～5:1)得到化合物44c。MS ESI m/z:309.3,311.3[M+H]+。Add compound 44b (3.58g, 20.81mmol) into a 100mL three-neck reaction flask, replace tetrahydrofuran (30mL) with nitrogen three times, cool the temperature to 0°C, add sodium hydrogen (1.39g, 34.68mmol, 60% purity), stir at 0°C for 0.5 hours, and add 4 , 6-dichloronicotinonitrile (3g, 17.34mmol), stirred at 25°C for 10 hours. After the reaction is completed, the reaction solution is quenched by adding saturated aqueous ammonium chloride solution (40 mL), extracted with ethyl acetate (50 mL × 2), and the organic phase is collected. The organic phase is washed with saturated brine (40 mL) and dried over anhydrous sodium sulfate. Filter and concentrate under reduced pressure to obtain the product. The product was purified by column chromatography (petroleum ether: ethyl acetate = 10:0 to 5:1) to obtain compound 44c. MS ESI m/z:309.3,311.3[M+H] + .
步骤3Step 3
在反应瓶中加入浓硫酸(0.5mL)，冰醋酸(0.5mL)，水(0.5mL)，60℃加入化合物44c(350mg,1.13mmol)，100℃搅拌2.5小时后，25℃搅拌12小时。反应液缓慢加入到冰水(20g)中淬灭，使用饱和碳酸氢钠水溶液调节pH＝6-8，使用乙酸乙酯萃取(30mL×3)，收集有机相，有机相用无水硫酸钠干燥，过滤，滤液减压浓缩得到粗品，再使用柱层析纯化(石油醚：乙酸乙酯＝10:0-3:1)得到化合物44d。MS ESI m/z:251.1,253.1[M+H]+ Concentrated sulfuric acid (0.5 mL), glacial acetic acid (0.5 mL), and water (0.5 mL) were added to the reaction flask. Compound 44c (350 mg, 1.13 mmol) was added at 60°C, stirred at 100°C for 2.5 hours, and then stirred at 25°C for 12 hours. The reaction solution was slowly added to ice water (20g) to quench, use saturated sodium bicarbonate aqueous solution to adjust pH=6-8, use ethyl acetate to extract (30mL×3), collect the organic phase, and dry the organic phase with anhydrous sodium sulfate , filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography (petroleum ether: ethyl acetate = 10:0-3:1) to obtain compound 44d. MS ESI m/z:251.1,253.1[M+H] +
步骤4Step 4
在反应瓶中加入化合物44d(50mg,199.42μmol)，二氯甲烷(6mL)，三乙胺(80.72mg,797.69μmol,111.03μL)，氮气置换三次降温至0℃滴加三氟甲磺酸酐(140.66mg,498.56μmol,82.26μL)，0℃搅拌1小时。该反应液中加入氯化铵(5mL)淬灭后使用二氯甲烷萃取(10mL×3)，无水硫酸钠干燥，过滤，减压浓缩得到产物44e。无后续纯化，直接用于下一步反应。Add compound 44d (50 mg, 199.42 μmol), dichloromethane (6 mL), and triethylamine (80.72 mg, 797.69 μmol, 111.03 μL) into the reaction bottle. Replace with nitrogen three times and cool to 0°C. Add trifluoromethanesulfonic anhydride ( 140.66 mg, 498.56 μmol, 82.26 μL), stir at 0°C for 1 hour. The reaction solution was quenched by adding ammonium chloride (5 mL), extracted with dichloromethane (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain product 44e. No subsequent purification was required and it was used directly in the next reaction.
步骤5Step 5
将异丙醇(3mL)，化合物44e(40mg,104.50μmol)，化合物28h(25.59mg,128.13μmol)加入长管中，开始搅拌；然后将N,N-二异丙基乙胺(81.03mg,626.98μmol,109.21μL)加入其中，升温至100℃，反应20小时。反应液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～15:85)，得到化合物44f。MS ESI m/z:395.9,397.9[M+H]+。Add isopropyl alcohol (3mL), compound 44e (40mg, 104.50μmol), and compound 28h (25.59mg, 128.13μmol) into a long tube and start stirring; then add N,N-diisopropylethylamine (81.03mg, 626.98 μmol, 109.21 μL) was added, the temperature was raised to 100°C, and the reaction was carried out for 20 hours. The reaction solution was concentrated under reduced pressure at 45°C. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~15:85) to obtain compound 44f. MS ESI m/z:395.9,397.9[M+H] + .
步骤6Step 6
将1,4-二氧六环(1.5mL)，化合物44f(11mg,27.78μmol)，化合物23f(6.29mg,27.78μmol)加入长管中，开始搅拌；氮气置换后，依次将乙酸钾(8.18mg,83.34μmol)，BrettPhos Pd G3(5.04mg,5.56μmol)加入其中，升温至100℃，反应2.5小时。向反应体系中加入15mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2次)；得到有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经薄层色谱分离(展开剂为乙酸乙酯：甲醇＝40:1)，得到化合物44。MS ESI m/z:586.6[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.35(s,1H),8.09(d,J＝5.6Hz,1H),7.53(s,1H),6.17(d,J＝5.6Hz,1H),4.91-4.77(m,1H),4.66-4.57(m,2H),4.55-4.48(m,1H),4.34-4.30(m,1H),3.88-3.78(m,1H),3.70(t,J＝7.6Hz,1H),3.64-3.59(m,2H),3.53(s,3H),3.40-3.35(m,1H),3.30-3.24(m,1H),3.11-3.06(m,1H),3.30-2.92(m,5H),2.32-2.25(m,4H),2.07-2.01(m,1H),1.89-1.82(m,1H),1.56(d,J＝6.0Hz,3H),1.01-0.99(m,6H)。Add 1,4-dioxane (1.5mL), compound 44f (11mg, 27.78μmol), and compound 23f (6.29mg, 27.78μmol) into a long tube and start stirring; after nitrogen replacement, add potassium acetate (8.18 mg, 83.34 μmol), BrettPhos Pd G3 (5.04 mg, 5.56 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 2.5 hours. After adding 15 mL of saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. The crude product was separated by thin layer chromatography (developing solvent: ethyl acetate: methanol = 40:1) to obtain compound 44. MS ESI m/z:586.6[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07 (s, 1H), 8.35 (s, 1H), 8.09 (d, J = 5.6Hz, 1H), 7.53 (s, 1H), 6.17 (d, J = 5.6 Hz,1H),4.91-4.77(m,1H),4.66-4.57(m,2H),4.55-4.48(m,1H),4.34-4.30(m,1H),3.88-3.78(m,1H), 3.70(t,J＝7.6Hz,1H),3.64-3.59(m,2H),3.53(s,3H),3.40-3.35(m,1H),3.30-3.24(m,1H),3.11-3.06( m,1H),3.30-2.92(m,5H),2.32-2.25(m,4H),2.07-2.01(m,1H),1.89-1.82(m,1H),1.56(d,J＝6.0Hz, 3H),1.01-0.99(m,6H).
实施例45
Example 45
步骤1step 1
在反应瓶中加入4-氨基-2-氯嘧啶(500mg,3.86mmol)，4-(二甲基氨基)哌啶(494.86mg,3.86mmol)，异丙醇(5mL)，三乙胺(2.34g,23.16mmol,3.22mL)氮气置换三次，100℃搅拌12hr。反应液进行减压浓缩得到粗品，再经柱层析(二氯甲烷：甲醇＝10:0～5:1)纯化得到化合物45a。1H NMR(400MHz,CD3OD)δppm7.73(d,J＝8.00Hz,1H),5.83(d,J＝5.88Hz,1H),4.74(d,J＝12.00Hz,2H),2.88-2.97(m,1H),2.76-2.86(m,2H),2.57(s,6H),1.99(d,J＝12.00Hz,2H),1.48(m,2H)。Add 4-amino-2-chloropyrimidine (500mg, 3.86mmol), 4-(dimethylamino)piperidine (494.86mg, 3.86mmol), isopropanol (5mL), and triethylamine (2.34 g, 23.16 mmol, 3.22 mL), replaced with nitrogen three times, and stirred at 100°C for 12 hr. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography (dichloromethane:methanol=10:0~5:1) to obtain compound 45a. 1H NMR (400MHz, CD3OD) δppm7.73(d,J＝8.00Hz,1H),5.83(d,J＝5.88Hz,1H),4.74(d,J＝12.00Hz,2H),2.88-2.97(m ,1H),2.76-2.86(m,2H),2.57(s,6H),1.99(d,J＝12.00Hz,2H),1.48(m,2H).
步骤2Step 2
在反应瓶中加入化合物41e(60mg,152.31μmol)，化合物45a(33.71mg,152.31μmol)，二氧六环(2mL)，氮气置换三次加入BrettPhos Pd G3(27.61mg,30.46μmol)，乙酸钾(44.84mg,456.93μmol)，100℃搅拌2小时。反应液冷却至室温后加入水(5mL)，使用乙酸乙酯萃取(10mL×3)，收集有机相，有机相使用无水硫酸钠干燥，过滤，减压浓缩得到产物粗品。粗品经制备高效液相色谱分离(色谱柱:Waters Xbridge Prep OBD C18 150×40mm×10μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:35-65％，8min)得到化合物45。MS ESI m/z:579.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.05(s,1H),8.66(s,1H),8.08(d,J＝5.63Hz,1H),7.41(d,J＝7.88Hz,1H),7.28(s,1H),6.46(d,J＝8.00Hz,1H),6.12(d,J＝4.00Hz,1H),4.94(d,J＝8.00Hz,2H),4.76(m,1H),4.42(t,J＝8.00Hz,2H),4.10(t,J＝7.07Hz,2H),3.53-3.65(m,1H),3.44-3.51(m,2H),3.08-3.13(m,2H),2.97(br t,J＝12.00Hz,2H),2.45-2.55(m,1H),2.36(s,6H),2.25(m,2H),1.95(d,J＝12.00Hz,2H),1.74-1.81(m,2H),1.52-1.61(m,2H),1.36(d,J＝4.00Hz,6H)。Add compound 41e (60mg, 152.31μmol), compound 45a (33.71mg, 152.31μmol), dioxane (2mL) to the reaction bottle, replace with nitrogen three times and add BrettPhos Pd G3 (27.61mg, 30.46μmol), potassium acetate ( 44.84 mg, 456.93 μmol), stir at 100°C for 2 hours. After the reaction solution was cooled to room temperature, water (5 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Waters 65%, 8 min) to obtain compound 45. MS ESI m/z:579.4[M+H]+. 1H NMR (400MHz, CDCl3) δppm 9.05 (s, 1H), 8.66 (s, 1H), 8.08 (d, J = 5.63Hz, 1H), 7.41 (d, J = 7.88Hz, 1H), 7.28 (s, 1H),6.46(d,J＝8.00Hz,1H),6.12(d,J＝4.00Hz,1H),4.94(d,J＝8.00Hz,2H),4.76(m,1H),4.42(t, J＝8.00Hz,2H),4.10(t,J＝7.07Hz,2H),3.53-3.65(m,1H),3.44-3.51(m,2H),3.08-3.13(m,2H),2.97(br t,J＝12.00Hz,2H),2.45-2.55(m,1H),2.36(s,6H),2.25(m,2H),1.95(d,J＝12.00Hz,2H),1.74-1.81(m ,2H),1.52-1.61(m,2H),1.36(d,J=4.00Hz,6H).
实施例46、47
Examples 46 and 47
步骤1step 1
将二氯甲烷(4mL)，四氢噻喃(0.2g,1.96mmol)加入拇指瓶中，开始搅拌；然后将温度降至0～5℃后，将间氯过氧苯甲酸(1.03g,5.09mmol,85％)分批次加入其中，升至室温20℃，反应2小时。向反应体系中加入10mL饱和亚硫酸钠溶液和10mL二氯甲烷后，分液；有机相用10mL饱和亚硫酸钠溶液洗涤，加入无水硫酸钠干燥后，过滤，滤液在40℃下进行减压浓缩。加入1.5mL甲基叔丁醚混合搅拌10min后，过滤，滤饼为产物。收集所有水相加入亚硫酸钠溶液至淀粉碘化钾试纸不变蓝后丢弃。得到化合物46a。1H NMR(400MHz,CDCl3)δppm 3.01-2.98(m,4H),2.14-2.08(m,4H),1.68-1.62(m,2H)。Add methylene chloride (4mL) and tetrahydrothiofuran (0.2g, 1.96mmol) into the thumb bottle and start stirring; then lower the temperature to 0~5°C, add m-chloroperoxybenzoic acid (1.03g, 5.09 mmol, 85%) was added in batches, raised to room temperature 20°C, and reacted for 2 hours. After adding 10 mL of saturated sodium sulfite solution and 10 mL of methylene chloride to the reaction system, the layers were separated; the organic phase was washed with 10 mL of saturated sodium sulfite solution, dried by adding anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40°C. Add 1.5 mL of methyl tert-butyl ether, mix and stir for 10 minutes, then filter, and the filter cake is the product. Collect all the water phase and add sodium sulfite solution until the starch potassium iodide test paper does not turn blue and then discard it. Compound 46a was obtained. 1H NMR (400MHz, CDCl3) δppm 3.01-2.98(m,4H),2.14-2.08(m,4H),1.68-1.62(m,2H).
步骤2Step 2
将四氢呋喃(60mL)，化合物46a(3g,22.36mmol)加入100mL三口瓶中，开始搅拌；然后将温度降至-65～-60℃后，将正丁基锂的正己烷溶液(2.5M,8.94mL)滴加到其中，反应1小时；然后将3-氧代氮杂环丁烷-1-羧酸叔丁酯(3.83g,22.36mmol)溶于四氢呋喃(60mL)中后，缓慢滴加到其中，继续反应2小时。反应液缓慢倒入120mL饱和氯化铵溶液中后，加入乙酸乙酯萃取(50mL×2)；得到的有机相用120mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到粗品。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100：0～30：70)，得到化合物46b。MS ESI m/z:579.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 4.51(d,J＝10.0Hz,1H),4.02(d,J＝10.0Hz,1H),3.96-3.87(m,2H),3.26-3.23(m,1H),3.15-3.11(m,1H),3.05-2.97(m,2H),2.21-2.08(m,4H),1.68-1.51(m,2H),1.44(s,9H)。Add tetrahydrofuran (60mL) and compound 46a (3g, 22.36mmol) into a 100mL three-necked flask and start stirring; then lower the temperature to -65~-60℃, add n-butyllithium n-hexane solution (2.5M, 8.94 mL) was added dropwise to it and reacted for 1 hour; then, 3-oxoazetidine-1-carboxylic acid tert-butyl ester (3.83g, 22.36mmol) was dissolved in tetrahydrofuran (60mL) and slowly added dropwise to Among them, the reaction was continued for 2 hours. The reaction solution was slowly poured into 120 mL of saturated ammonium chloride solution, and then extracted with ethyl acetate (50 mL Concentrate under reduced pressure to obtain crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~30:70) to obtain compound 46b. MS ESI m/z:579.4[M+H]+. 1H NMR(400MHz, CDCl3)δppm 4.51(d,J＝10.0Hz,1H),4.02(d,J＝10.0Hz,1H),3.96-3.87(m,2H),3.26-3.23(m,1H), 3.15-3.11(m,1H),3.05-2.97(m,2H),2.21-2.08(m,4H),1.68-1.51(m,2H),1.44(s,9H).
步骤3Step 3
将二氯甲烷(50mL)，化合物46b(2.5g,8.19mmol)加入100mL三口瓶中开始搅拌；氮气置换后，将温度降至-20℃，然后将二乙氨基三氟化硫(1.52g,9.41mmol,1.24mL)滴加到其中，反应2小时。向反应体系中30mL饱和碳酸氢钠溶液后，分液；得到的有机相依次用饱和碳酸氢钠溶液(30mL)，饱和食盐水洗涤后，加入无水硫酸钠干燥后，过滤，滤液在40℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100：0～30：70)，得到化合物46c。MS ESI m/z:231.9[M+H]+。Add dichloromethane (50 mL) and compound 46b (2.5 g, 8.19 mmol) into a 100 mL three-necked flask and start stirring; after nitrogen replacement, lower the temperature to -20°C, and then add diethylamino sulfur trifluoride (1.52 g, 9.41 mmol, 1.24 mL) was added dropwise to it, and the reaction was carried out for 2 hours. Add 30 mL of saturated sodium bicarbonate solution to the reaction system and separate the layers; the obtained organic phase was washed with saturated sodium bicarbonate solution (30 mL) and saturated brine, and then dried with anhydrous sodium sulfate and filtered. The filtrate was heated at 40°C. Concentrate under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~30:70) to obtain compound 46c. MS ESI m/z:231.9[M+H]+.
步骤4Step 4
将乙酸乙酯(40mL)，化合物46c(2.46g,8.56mmol)，钯/碳(910.98mg,856.03μmol,10％)加入 氢化瓶，室温20℃，氢气15psi反应20小时。反应液直接过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～30:70)，得到化合物46d。1H NMR(400MHz,CDCl3)δppm 4.16-4.05(m,3H),3.76(brs,1H),3.11-2.90(m,4H),2.10-2.00(m,3H),1.94-1.59(m,3H),1.43-1.42(m,9H)。Ethyl acetate (40 mL), compound 46c (2.46 g, 8.56 mmol), palladium on carbon (910.98 mg, 856.03 μmol, 10%) were added Hydrogenation bottle, room temperature 20℃, hydrogen 15psi, react for 20 hours. The reaction solution was directly filtered, and the filtrate was concentrated under reduced pressure at 45°C. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~30:70) to obtain compound 46d. 1H NMR (400MHz, CDCl3) δppm 4.16-4.05(m,3H),3.76(brs,1H),3.11-2.90(m,4H),2.10-2.00(m,3H),1.94-1.59(m,3H) ,1.43-1.42(m,9H).
步骤5Step 5
将乙酸乙酯(2mL)，化合物46d(0.4g,1.38mmol)加入50mL单口瓶中，开始搅拌，然后将盐酸/乙酸乙酯(4M,15mL)加入其中，室温20℃，反应2小时。反应液直接过滤，得到化合物46e的盐酸盐。Add ethyl acetate (2 mL) and compound 46d (0.4 g, 1.38 mmol) into a 50 mL single-neck bottle, start stirring, and then add hydrochloric acid/ethyl acetate (4 M, 15 mL), and react at room temperature 20°C for 2 hours. The reaction solution was filtered directly to obtain the hydrochloride salt of compound 46e.
步骤6Step 6
将二氧六环(10mL)，化合物46e的盐酸盐(278.42mg,1.23mmol)，化合物15g(270mg,948.77μmol)加入50mL单口瓶中，开始搅拌；氮气置换后，依次将碳酸铯(1.24g,3.80mmol)，甲烷磺酸[9,9-二甲基-4,5-双(二苯基膦基)氧杂蒽](2-甲胺基-1,1-联苯-2-基)钯(II)(136.96mg,142.32μmol)加入其中，升温至100℃，反应4小时。向反应体系中加入20mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×2)；得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～25:75)。得到化合物46f。MS ESI m/z:393.2[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.13(s,1H),7.84(s,1H),7.47(d,J＝8.0Hz,1H),6.53(d,J＝8.0Hz,1H),4.43(t,J＝7.6Hz,2H),4.23(t,J＝7.2Hz,1H),3.96-3.93(m,1H),3.50-3.36(m,2H),3.21-3.12(m,2H),3.00-2.92(m,1H),2.13-2.06(m,3H),1.97-1.79(m,2H),1.53-1.43(m,1H),1.34(d,J＝6.8,6H)。Add dioxane (10 mL), the hydrochloride of compound 46e (278.42 mg, 1.23 mmol), and compound 15 g (270 mg, 948.77 μmol) into a 50 mL single-mouth bottle, and start stirring; after nitrogen replacement, add cesium carbonate (1.24 g, 3.80mmol), methanesulfonic acid [9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene](2-methylamino-1,1-biphenyl-2- (136.96 mg, 142.32 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 4 hours. After adding 20 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:0~25:75). Compound 46f was obtained. MS ESI m/z:393.2[M+H]+. 1 H NMR (400MHz, CDCl3) δppm 9.13 (s, 1H), 7.84 (s, 1H), 7.47 (d, J = 8.0Hz, 1H), 6.53 (d, J = 8.0Hz, 1H), 4.43 (t ,J＝7.6Hz,2H),4.23(t,J＝7.2Hz,1H),3.96-3.93(m,1H),3.50-3.36(m,2H),3.21-3.12(m,2H),3.00- 2.92(m,1H),2.13-2.06(m,3H),1.97-1.79(m,2H),1.53-1.43(m,1H),1.34(d,J=6.8,6H).
步骤7Step 7
将二氧六环(10mL)，化合物46f(0.1g,254.49μmol)，化合物23f(51.82mg,229.04μmol)加入长管中，开始搅拌；氮气置换后，依次将碳酸铯(248.75mg,763.47μmol)，甲磺酸(2-二环己基膦基-3,6-二甲氧基-2,4,6-三异丙基-1,1-联苯)(2-氨基-1,1-联苯-2-基)钯(II)(34.60mg,38.17μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入20mL饱和氯化铵溶液后，加入乙酸乙酯萃取(10mL×3)；得到的有机相用20mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品经硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～10:90)，再经过SFC拆分(色谱柱：DAICEL CHIRALCEL OX(250mm×30mm，10μm)；流动相：A(超临界CO2)和B(异丙醇，含0.1％氨水)；梯度：B％＝60％-60％，35min)得到化合物46和化合物47。Add dioxane (10 mL), compound 46f (0.1g, 254.49 μmol), and compound 23f (51.82 mg, 229.04 μmol) into the long tube and start stirring; after nitrogen replacement, add cesium carbonate (248.75 mg, 763.47 μmol) in sequence. ), methanesulfonic acid (2-dicyclohexylphosphino-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl) (2-amino-1,1- Biphenyl-2-yl)palladium (II) (34.60 mg, 38.17 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 3 hours. After adding 20 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (10 mL Concentrate under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:0~10:90), and then separated by SFC (chromatographic column: DAICEL CHIRALCEL OX (250mm×30mm, 10μm); mobile phase: A (ultra Critical CO 2 ) and B (isopropanol, containing 0.1% ammonia); gradient: B% = 60%-60%, 35min) to obtain compound 46 and compound 47.
化合物46：MS ESI m/z:583.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.57(s,1H),8.08(d,J＝5.6Hz,1H),7.41(d,J＝8.0Hz,1H),7.32(s,1H),6.42(d,J＝8.0Hz,1H),6.19(d,J＝5.6Hz,1H),4.91-4.78(m,1H),4.64-4.57(m,1H),4.47-4.39(m,2H),4.34-4.31(m,1H),4.23(d,J＝7.2Hz,1H),3.95-3.92(m,1H),3.86-3.76(m,1H),3.66-3.56(m,3H),3.52(s,3H),3.44-3.35(m,1H),3.21-3.12(m,2H),3.00-2.92(m,1H),2.14-2.03(m,4H),1.97-1.84(m,3H),1.51-1.46(m,1H),1.37(d,J＝5.6Hz,6H)。SFC分析检测(色谱柱：Chiralcel OX-3，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(异丙醇，含0.1％异丙胺)；梯度：B％＝50～50％，6min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝2.261min，ee：100％。Compound 46: MS ESI m/z: 583.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07 (s, 1H), 8.57 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.41 (d, J = 8.0Hz, 1H), 7.32 ( s,1H),6.42(d,J＝8.0Hz,1H),6.19(d,J＝5.6Hz,1H),4.91-4.78(m,1H),4.64-4.57(m,1H),4.47-4.39 (m,2H),4.34-4.31(m,1H),4.23(d,J＝7.2Hz,1H),3.95-3.92(m,1H),3.86-3.76(m,1H),3.66-3.56(m ,3H),3.52(s,3H),3.44-3.35(m,1H),3.21-3.12(m,2H),3.00-2.92(m,1H),2.14-2.03(m,4H),1.97-1.84 (m,3H),1.51-1.46(m,1H),1.37(d,J=5.6Hz,6H). SFC analysis and detection (chromatographic column: Chiralcel OX-3, 3μm, 0.46cm id×5cm L; mobile phase: A (supercritical CO 2 ) and B (isopropanol, containing 0.1% isopropylamine); gradient: B%＝ 50～50%, 6min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=2.261min, ee: 100%.
化合物47：MS ESI m/z:583.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.07(s,1H),8.57(s,1H),8.08(d,J＝5.6Hz,1H),7.41(d,J＝8.0Hz,1H),7.35(s,1H),6.42(d,J＝8.0Hz,1H),6.19(d,J＝5.6Hz,1H),4.91-4.78(m,1H),4.64-4.57(m,1H),4.47-4.39(m,2H),4.34-4.30(m,1H),4.24-4.21(m,1H),3.95-3.92(m,1H),3.86-3.76(m,1H),3.66-3.56(m,3H),3.52(s,3H),3.44-3.35(m,1H),3.21-3.11(m,2H),3.00-2.92(m,1H),2.13-2.03(m,4H),1.97-1.81(m,3H),1.54-1.44(m,1H),1.36(dd,J＝6.8and 2.4Hz,6H)。SFC分析检测(色谱柱：Lux Cellulose-2，3μm，0.46cm id×5cm L；流动相：A(超临界CO2)和B(乙醇，含0.1％异丙胺)；梯度：B％＝50～50％，4min；流速:4.0mL/min；波长:220nm；压力:1800psi，Rt＝3.348min，ee：100％。Compound 47: MS ESI m/z: 583.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.07 (s, 1H), 8.57 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.41 (d, J = 8.0Hz, 1H), 7.35 ( s,1H),6.42(d,J＝8.0Hz,1H),6.19(d,J＝5.6Hz,1H),4.91-4.78(m,1H),4.64-4.57(m,1H),4.47-4.39 (m,2H),4.34-4.30(m,1H),4.24-4.21(m,1H),3.95-3.92(m,1H),3.86-3.76(m,1H),3.66-3.56(m,3H) ,3.52(s,3H),3.44-3.35(m,1H),3.21-3.11(m,2H),3.00-2.92(m,1H),2.13-2.03(m,4H),1.97-1.81(m, 3H),1.54-1.44(m,1H),1.36(dd,J＝6.8and 2.4Hz,6H). SFC analysis and detection (chromatographic column: Lux Cellulose-2, 3μm, 0.46cm id×5cm L; mobile phase: A (supercritical CO 2 ) and B (ethanol, containing 0.1% isopropylamine); gradient: B%=50~ 50%, 4min; flow rate: 4.0mL/min; wavelength: 220nm; pressure: 1800psi, Rt=3.348min, ee: 100%.
实施例48
Example 48
步骤1step 1
在反应瓶中加入4-氨基-2-氯嘧啶(200mg,1.54mmol)，化合物48a(306.08mg,1.54mmol)，异丙醇(6mL)，三乙胺(937.32mg,9.26mmol,1.29mL)100℃搅拌12hr。反应液在45℃下进行减压浓缩得到粗品，粗品经硅胶柱层析纯化(二氯甲烷：甲醇＝10:0～10:1)。得到化合物48b。1H NMR(400MHz,CDCl3)δppm 7.98(d,J＝8.00Hz,1H),5.83(d,J＝4.00Hz,1H),4.59(s,2H),4.24(s,2H),3.59(d,J＝12.00Hz,2H),3.24(d,J＝12.00Hz,1H),2.61(m,1H),1.46(d,J＝8.00Hz,2H),1.39(s,9H)。Add 4-amino-2-chloropyrimidine (200mg, 1.54mmol), compound 48a (306.08mg, 1.54mmol), isopropyl alcohol (6mL), and triethylamine (937.32mg, 9.26mmol, 1.29mL) into the reaction bottle. Stir at 100°C for 12 hours. The reaction solution was concentrated under reduced pressure at 45°C to obtain a crude product, which was purified by silica gel column chromatography (dichloromethane: methanol = 10:0 to 10:1). Compound 48b was obtained. 1 H NMR (400MHz, CDCl 3 ) δppm 7.98 (d, J = 8.00Hz, 1H), 5.83 (d, J = 4.00Hz, 1H), 4.59 (s, 2H), 4.24 (s, 2H), 3.59 ( d, J＝12.00Hz, 2H), 3.24 (d, J＝12.00Hz, 1H), 2.61 (m, 1H), 1.46 (d, J＝8.00Hz, 2H), 1.39 (s, 9H).
步骤2Step 2
在反应瓶中加入化合物41e(80mg,203.08μmol)，化合物48b(59.17mg,203.08μmol)，二氧六环(6mL)，氮气置换三次加入乙酸钾(59.79mg,609.25μmol)，BrettPhos Pd G3(36.82mg,40.62μmol)，100℃搅拌2小时。反应液中加入水(10mL)使用乙酸乙酯萃取(15mL×3)，收集有机相，有机相使用饱和实验室洗涤(20mL)，无水硫酸钠干燥，过滤，减压浓缩得到产物。产物通过柱层析纯化(二氯甲烷：甲醇＝10:0～10:1)纯化得到化合物48c。MS ESI m/z:649.3[M+H]+。Add compound 41e (80 mg, 203.08 μmol), compound 48b (59.17 mg, 203.08 μmol), dioxane (6 mL) to the reaction bottle, replace with nitrogen three times and add potassium acetate (59.79 mg, 609.25 μmol), BrettPhos Pd G3 ( 36.82 mg, 40.62 μmol), stir at 100°C for 2 hours. Water (10 mL) was added to the reaction solution and extracted with ethyl acetate (15 mL × 3). The organic phase was collected, washed with saturated laboratory (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the product. The product was purified by column chromatography (dichloromethane:methanol=10:0~10:1) to obtain compound 48c. MS ESI m/z:649.3[M+H] + .
步骤3Step 3
在反应瓶中加入化合物48c(100mg,92.48μmol)，二氯甲烷(5mL)，三氟乙酸(1mL)，25℃搅拌1小时。反应液使用1N氢氧化钠水溶液调节pH＝8-9，使用二氯甲烷萃取(10mL×3)，无水硫酸钠干燥，过滤，滤液减压浓缩得到化合物48d。MS ESI m/z:549.4[M+H]+。Compound 48c (100 mg, 92.48 μmol), dichloromethane (5 mL), and trifluoroacetic acid (1 mL) were added to the reaction flask, and stirred at 25°C for 1 hour. The reaction solution was adjusted to pH=8-9 using 1N sodium hydroxide aqueous solution, extracted with dichloromethane (10 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 48d. MS ESI m/z:549.4[M+H] + .
步骤4Step 4
在反应瓶中加入化合物48d(120mg,218.70μmol)，甲醇(10mL)，甲醛(35.50mg,437.40μmol,32.56μL,37％)，醋酸硼氢化钠(278.11mg,1.31mmol)，冰醋酸(39.40mg,656.09μmol,37.56μL)，25℃搅拌1小时。反应液使用1N氢氧化钠水溶液调节pH＝8-9，使用二氯甲烷萃取(10mL×3)，收集有机相，有机相使用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品中加入甲醇(6mL)搅拌10min后过滤，滤饼减压浓缩得到产物。得到化合物48。MS ESI m/z:563.4[M+H]+。1H NMR(400 MHz,CDCl3)δppm 9.06(s,1H),8.88(s,1H),8.14(d,J＝8.00Hz,1H),7.41(d,J＝8.00Hz,1H),7.35(s,1H),6.46(d,J＝8.00Hz,1H),6.18(d,J＝8.00Hz,1H),4.76(t,J＝8.00Hz,1H),4.42(t,J＝8.00Hz,2H),4.11(m,3H),3.84-3.97(m,2H),3.74(m,3H),3.52-3.63(m,1H),3.43-3.51(m,2H),3.07-3.15(m,2H),2.66-2.77(m,1H),2.19-2.33(m,5H),1.77(s,2H),1.67(d,J＝8.00Hz,1H),1.34(d,J＝8.00Hz,6H)。Add compound 48d (120 mg, 218.70 μmol), methanol (10 mL), formaldehyde (35.50 mg, 437.40 μmol, 32.56 μL, 37%), sodium acetate borohydride (278.11 mg, 1.31 mmol), and glacial acetic acid (39.40) into the reaction bottle. mg, 656.09 μmol, 37.56 μL), stir at 25°C for 1 hour. The reaction solution was adjusted to pH=8-9 using 1N sodium hydroxide aqueous solution, extracted with dichloromethane (10mL×3), and the organic phase was collected. The organic phase was washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and reduced to Concentrate under pressure to obtain crude product. Methanol (6 mL) was added to the crude product and stirred for 10 min, then filtered, and the filter cake was concentrated under reduced pressure to obtain the product. Compound 48 was obtained. MS ESI m/z:563.4[M+H] + . 1 H NMR(400 MHz, CDCl 3 )δppm 9.06(s,1H),8.88(s,1H),8.14(d,J＝8.00Hz,1H),7.41(d,J＝8.00Hz,1H),7.35(s,1H) ,6.46(d,J＝8.00Hz,1H),6.18(d,J＝8.00Hz,1H),4.76(t,J＝8.00Hz,1H),4.42(t,J＝8.00Hz,2H),4.11 (m,3H),3.84-3.97(m,2H),3.74(m,3H),3.52-3.63(m,1H),3.43-3.51(m,2H),3.07-3.15(m,2H),2.66 -2.77(m,1H),2.19-2.33(m,5H),1.77(s,2H),1.67(d,J＝8.00Hz,1H),1.34(d,J＝8.00Hz,6H).
实施例49
Example 49
步骤1step 1
将异丙醇(7mL)，4-氨基-2-氯嘧啶(0.2g,1.54mmol)，化合物49a(491.61mg,2.32mmol)加入长管中，开始搅拌；然后将三乙胺(781.10mg,7.72mmol,1.07mL)加入其中，升温至100℃反应20小时。向反应体系中加入20mL水后，加入乙酸乙酯萃取(10mL×2)；得到的有机相用饱和氯化铵溶液洗涤(15mL×2)后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得到化合物49b。MS ESI m/z:306.3[M+H]+。1H NMR(400MHz,CDCl3)δppm 7.93(d,J＝5.6Hz,1H),5.78(d,J＝5.6Hz,1H),4.62(s,2H),3.81-3.73(m,2H),3.65-3.59(m,2H),3.50-3.46(m,2H),3.35-3.23(m,2H),2.90-2.98(m,2H),1.45(s,9H)。Add isopropanol (7mL), 4-amino-2-chloropyrimidine (0.2g, 1.54mmol), and compound 49a (491.61mg, 2.32mmol) into a long tube and start stirring; then triethylamine (781.10mg, 7.72mmol, 1.07mL) was added, and the temperature was raised to 100°C for reaction for 20 hours. After adding 20 mL of water to the reaction system, add ethyl acetate for extraction (10 mL × 2); wash the obtained organic phase with saturated ammonium chloride solution (15 mL × 2), add anhydrous sodium sulfate, dry, filter, and the filtrate is dried at 45 Concentrate under reduced pressure at ℃ to obtain compound 49b. MS ESI m/z:306.3[M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.93 (d, J＝5.6Hz, 1H), 5.78 (d, J＝5.6Hz, 1H), 4.62 (s, 2H), 3.81-3.73 (m, 2H), 3.65 -3.59(m,2H),3.50-3.46(m,2H),3.35-3.23(m,2H),2.90-2.98(m,2H),1.45(s,9H).
步骤2Step 2
将1,4-二氧六环(10mL)，化合物19e(165.78mg,734.42μmol)，化合物15g(190mg,667.65μmol)加入长管中，开始搅拌；氮气置换后路，依次将碳酸铯(652.60mg,2.00mmol)，甲磺酸(4,5-双二苯基膦-9,9-二甲基氧杂蒽)(2'-甲胺基-1,1'-联苯-2-基)钯(II)(96.38mg,100.15μmol)加入其中，升温至100℃，反应3小时。向反应体系中加入20mL饱和氯化铵溶液后，加入乙酸乙酯萃取(15mL×2)；得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩。粗品通过硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～10:90)，得到消旋化合物，再经过SFC进行拆分(色谱柱：DAICEL CHIRALCEL OZ(250mm×25mm,10μm)；流动相：A(超临界CO2)和B(甲醇)；梯度：B％＝55％-55％，isocratic elution mode,rt＝2.054min)，得到化合物49c。MS ESI m/z:393.1,395.1[M+H]+。Add 1,4-dioxane (10mL), compound 19e (165.78mg, 734.42μmol), and compound 15g (190mg, 667.65μmol) into a long tube and start stirring; replace the rear path with nitrogen and add cesium carbonate (652.60 mg, 2.00mmol), methanesulfonic acid (4,5-bisdiphenylphosphine-9,9-dimethylxanthene) (2'-methylamino-1,1'-biphenyl-2-yl ) Palladium (II) (96.38 mg, 100.15 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 3 hours. After adding 20 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (15 mL Concentrate under reduced pressure. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0 ~ 10:90) to obtain the racemic compound, which was then separated by SFC (chromatographic column: DAICEL CHIRALCEL OZ (250mm× 25mm, 10μm); mobile phase: A (supercritical CO 2 ) and B (methanol); gradient: B%=55%-55%, isocratic elution mode, rt=2.054min) to obtain compound 49c. MS ESI m/z:393.1,395.1[M+H]+.
步骤3Step 3
将1,4-二氧六环(3mL)，化合物49c(40.00mg,101.80μmol)，化合物49b(37.30mg,122.16μmol)加入长管中，开始搅拌；氮气置换后，依次将碳酸铯(132.67mg,407.19μmol)，BrettPhos Pd G3(18.46mg,20.36μmol)加入其中，升温至100℃，反应4小时。向反应体系中加入10mL饱和氯化铵溶液后，加入乙酸乙酯萃取(5mL×3)；得到的有机相用10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进 行减压浓缩。粗品通过硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～10:90)，得到化合物49d。MS ESI m/z:662.3[M+H]+。Add 1,4-dioxane (3mL), compound 49c (40.00mg, 101.80μmol), and compound 49b (37.30mg, 122.16μmol) into a long tube and start stirring; after nitrogen replacement, add cesium carbonate (132.67 mg, 407.19 μmol), BrettPhos Pd G3 (18.46 mg, 20.36 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 4 hours. After adding 10 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (5 mL Enter Concentrate under reduced pressure. The crude product was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0 to 10:90) to obtain compound 49d. MS ESI m/z:662.3[M+H]+.
步骤4Step 4
将二氯甲烷(1.5mL)，化合物49d(15.00mg,22.66μmol)加入拇指瓶中，开始搅拌；然后将三氟乙酸(460.50mg,4.04mmol,0.3mL)滴加到其中，室温15℃，反应1小时。反应液在40℃下进行减压浓缩后，加入10mL二氯甲烷和10mL饱和碳酸氢钠溶液后，分液；有机相依次用10mL饱和碳酸氢钠溶液，10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在40℃下进行减压浓缩。得到化合物49e。Add dichloromethane (1.5 mL) and compound 49d (15.00 mg, 22.66 μmol) into the thumb bottle and start stirring; then add trifluoroacetic acid (460.50 mg, 4.04 mmol, 0.3 mL) dropwise into it, at room temperature 15°C. Reaction takes 1 hour. After the reaction solution was concentrated under reduced pressure at 40°C, 10 mL of methylene chloride and 10 mL of saturated sodium bicarbonate solution were added, and the liquids were separated; the organic phase was washed with 10 mL of saturated sodium bicarbonate solution and 10 mL of saturated brine, and then anhydrous was added. Dry over sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 40°C. Compound 49e was obtained.
步骤5Step 5
将甲醇(1.5mL)，化合物49e(8mg,14.24μmol),甲醛溶液(11.56mg,142.41μmol,10.60μL,37％)加入拇指瓶中，开始搅拌，室温20℃，反应0.5小时，然后将三乙酰氧基硼氢化钠(12.07mg,56.97μmol,4eq)加到其中，继续反应0.5小时。向反应体系中加入10mL水后，加入0.5mL饱和碳酸钠溶液，加入二氯甲烷萃取(10mL×2)；得到的有机相依次用10mL饱和碳酸氢钠溶液，10mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在40℃下进行减压浓缩；得到化合物49。MS ESI m/z:576.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.06(s,1H),8.85(s,1H),8.06(d,J＝5.6Hz,1H),7.53(s,1H),7.39(d,J＝8.0Hz,1H),6.48(d,J＝8.0Hz,1H),6.10(d,J＝6.0Hz,1H),4.69(t,J＝7.6Hz,1H),4.24-4.21(m,1H),3.93-3.85(m,2H),3.82-3.78(m,1H),3.74-3.71(m,1H),3.66-3.59(m,1H),3.33-3.26(m,1H),3.20-3.14(m,1H),3.07-2.99(m,3H),2.94-2.84(m,2H),2.80-2.72(m,1H),2.52-2.43(m,3H),2.40(s,2H),2.35-2.26(m,1H),2.20-2.10(m,2H),1.96-1.86(m,1H),1.60-1.71(m,1H),1.45(d,J＝6.0Hz,3H),1.37-1.34(m,6H)。Add methanol (1.5 mL), compound 49e (8 mg, 14.24 μmol), and formaldehyde solution (11.56 mg, 142.41 μmol, 10.60 μL, 37%) into the thumb bottle, start stirring, and react at room temperature 20°C for 0.5 hours, and then add the three Sodium acetoxyborohydride (12.07 mg, 56.97 μmol, 4 eq) was added to it, and the reaction was continued for 0.5 hours. After adding 10mL of water to the reaction system, add 0.5mL of saturated sodium carbonate solution, and add dichloromethane for extraction (10mL×2); the obtained organic phase was washed with 10mL of saturated sodium bicarbonate solution and 10mL of saturated brine, and then added Dry over sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 40°C to obtain compound 49. MS ESI m/z:576.4[M+H] + . 1H NMR (400MHz, CDCl3) δppm 9.06 (s, 1H), 8.85 (s, 1H), 8.06 (d, J = 5.6Hz, 1H), 7.53 (s, 1H), 7.39 (d, J = 8.0Hz, 1H),6.48(d,J＝8.0Hz,1H),6.10(d,J＝6.0Hz,1H),4.69(t,J＝7.6Hz,1H),4.24-4.21(m,1H),3.93- 3.85(m,2H),3.82-3.78(m,1H),3.74-3.71(m,1H),3.66-3.59(m,1H),3.33-3.26(m,1H),3.20-3.14(m,1H ),3.07-2.99(m,3H),2.94-2.84(m,2H),2.80-2.72(m,1H),2.52-2.43(m,3H),2.40(s,2H),2.35-2.26(m ,1H),2.20-2.10(m,2H),1.96-1.86(m,1H),1.60-1.71(m,1H),1.45(d,J＝6.0Hz,3H),1.37-1.34(m,6H ).
实施例50
Example 50
步骤1step 1
在反应瓶中加入化合物41e(80mg,203.08μmol)，化合物49b(62.02mg,203.08μmol)，二氧六环(5mL)氮气置换三次加入碳酸铯(198.50mg,609.25μmol)，BrettPhos Pd G3(36.82mg,40.62μmol)，100℃搅拌2小时。反应液中加入水(10mL)使用乙酸乙酯萃取(15mL×3)，收集有机相，有机相使用饱和实验室洗涤(20mL)，无水硫酸钠干燥，过滤，减压浓缩得到产物。产物通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物50a。MS ESI m/z:663.5[M+H]+。Add compound 41e (80 mg, 203.08 μmol) and compound 49b (62.02 mg, 203.08 μmol) to the reaction bottle. Dioxane (5 mL) is replaced with nitrogen three times and cesium carbonate (198.50 mg, 609.25 μmol) is added. BrettPhos Pd G3 (36.82 mg, 40.62 μmol), stir at 100°C for 2 hours. Water (10 mL) was added to the reaction solution and extracted with ethyl acetate (15 mL × 3). The organic phase was collected, washed with saturated laboratory (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the product. The product was purified by column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 50a. MS ESI m/z:663.5[M+H] + .
步骤2Step 2
在反应瓶中加入化合物50a(50mg,75.43μmol)，二氯甲烷(5mL)，三氟乙酸(1mL)，25℃搅拌1小时。反应液使用1N氢氧化钠水溶液调节pH＝8-9，使用二氯甲烷萃取(5mL×3)，无水硫酸钠干燥，过滤，减压浓缩得到化合物50b。直接用于下一步反应。MS ESI m/z:563.4[M+H]+。Compound 50a (50 mg, 75.43 μmol), dichloromethane (5 mL), and trifluoroacetic acid (1 mL) were added to the reaction flask, and stirred at 25°C for 1 hour. The reaction solution was adjusted to pH=8-9 using 1N sodium hydroxide aqueous solution, extracted with dichloromethane (5 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 50b. used directly for the next reaction. MS ESI m/z:563.4[M+H] + .
步骤3Step 3
在反应瓶中加入化合物50b(50mg,88.85μmol)，甲醇(10mL)，甲醛(14.42mg,177.71μmol,13.23μL,37％)，醋酸硼氢化钠(112.99mg,533.12μmol)，冰醋酸(16.01mg,266.56μmol,15.26μL)，25℃搅拌1小时。反应液中使用1N氢氧化钠水溶液调节pH＝8-9，使用二氯甲烷萃取(20mL×2)，饱和食盐水洗涤(10mL)， 无水硫酸钠干燥，过滤，减压浓缩得到粗品。粗品经制备高效液相色谱分离(色谱柱:Phenomenex Gemini-NX 80×40mm×3μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:25-55％，8min)得到化合物50。MS ESI m/z:577.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.05(s,1H),8.85(s,1H),8.07(d,J＝4.00Hz,1H),7.41(d,J＝8.00Hz,1H),7.30(s,1H),6.45(d,J＝8.00Hz,1H),6.09(d,J＝4.003Hz,1H),4.76(m,1H),4.42(t,J＝8.00Hz,2H),4.10(t,J＝8.00Hz,2H),3.87-3.96(m,2H),3.58-3.76(m,3H),3.44-3.52(m,2H),3.08-3.14(m,2H),3.03(s,2H),2.80(s,2H),2.51(m,2H),2.37(s,3H)2.25(m,2H),1.73-1.80(m,2H),1.36(d,J＝8.00Hz,6H)。Add compound 50b (50 mg, 88.85 μmol), methanol (10 mL), formaldehyde (14.42 mg, 177.71 μmol, 13.23 μL, 37%), sodium acetate borohydride (112.99 mg, 533.12 μmol), and glacial acetic acid (16.01) into the reaction bottle. mg, 266.56 μmol, 15.26 μL), stir at 25°C for 1 hour. Use 1N sodium hydroxide aqueous solution to adjust pH=8-9 in the reaction solution, extract with dichloromethane (20mL×2), and wash with saturated brine (10mL). Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain crude product. The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX 80×40mm×3μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% ammonia bicarbonate); gradient: B%: 25-55 %, 8 min) to obtain compound 50. MS ESI m/z:577.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.05 (s, 1H), 8.85 (s, 1H), 8.07 (d, J = 4.00Hz, 1H), 7.41 (d, J = 8.00Hz, 1H), 7.30 ( s,1H),6.45(d,J＝8.00Hz,1H),6.09(d,J＝4.003Hz,1H),4.76(m,1H),4.42(t,J＝8.00Hz,2H),4.10( t,J＝8.00Hz,2H),3.87-3.96(m,2H),3.58-3.76(m,3H),3.44-3.52(m,2H),3.08-3.14(m,2H),3.03(s, 2H),2.80(s,2H),2.51(m,2H),2.37(s,3H)2.25(m,2H),1.73-1.80(m,2H),1.36(d,J＝8.00Hz,6H) .
实施例51
Example 51
步骤1step 1
将异丙醇(7mL)，4-氨基-2-氯嘧啶(0.2g,1.54mmol)，化合物51a(491.61mg,2.32mmol)，加入长管中，开始搅拌；然后将三乙胺(781.10mg,7.72mmol,1.07mL)加入其中，升温至100℃，反应20小时，补加4-氨基-2-氯嘧啶(150mg)继续反应20小时。向反应体系中加入20mL水后，加入乙酸乙酯萃取(10mL×2)；得到的有机相用饱和氯化铵溶液洗涤(15mL×2)后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩，得到粗品；粗品与1.5mL乙酸乙酯混合搅拌10min后，过滤，滤液在45℃下进行减压浓缩，得化合物51b。MS ESI m/z:306.3[M+H]+。1H NMR(400MHz,CDCl3)δppm 7.93(d,J＝5.6Hz,1H),5.78(d,J＝5.6Hz,1H),4.71-4.67(m,2H),3.86-3.68(m,2H),3.20-3.08(m,2H),1.97-1.95(m,2H),1.82-1.76(m,2H),1.48(s,9H)。Add isopropanol (7mL), 4-amino-2-chloropyrimidine (0.2g, 1.54mmol), and compound 51a (491.61mg, 2.32mmol) into a long tube and start stirring; then add triethylamine (781.10mg , 7.72mmol, 1.07mL) was added, the temperature was raised to 100°C, and the reaction was carried out for 20 hours. 4-amino-2-chloropyrimidine (150mg) was added to continue the reaction for 20 hours. After adding 20 mL of water to the reaction system, add ethyl acetate for extraction (10 mL × 2); wash the obtained organic phase with saturated ammonium chloride solution (15 mL × 2), add anhydrous sodium sulfate, dry, filter, and the filtrate is dried at 45 Concentrate under reduced pressure at 45°C to obtain a crude product; the crude product was mixed with 1.5 mL of ethyl acetate and stirred for 10 min, then filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain compound 51b. MS ESI m/z:306.3[M+H] + . 1H NMR (400MHz, CDCl3) δppm 7.93 (d, J＝5.6Hz, 1H), 5.78 (d, J＝5.6Hz, 1H), 4.71-4.67 (m, 2H), 3.86-3.68 (m, 2H), 3.20-3.08(m,2H),1.97-1.95(m,2H),1.82-1.76(m,2H),1.48(s,9H).
步骤2Step 2
在反应瓶中加入化合物41e(80mg,203.08μmol)，化合物51b(62.02mg,203.08μmol)，二氧六环(5mL)，氮气置换三次加入碳酸铯(198.50mg,609.25μmol)，BrettPhos Pd G3(36.82mg,40.62μmol)，100℃搅拌2小时。反应液中加入水(10mL)使用乙酸乙酯萃取(15mL×3)，收集有机相，有机相使用饱和食盐水洗涤(20mL)，无水硫酸钠干燥，过滤，滤液减压浓缩得到粗产物。再通过柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)纯化得到化合物51c。MS ESI m/z:663.5[M+H]+。Add compound 41e (80 mg, 203.08 μmol), compound 51b (62.02 mg, 203.08 μmol), dioxane (5 mL) to the reaction bottle, replace with nitrogen three times and add cesium carbonate (198.50 mg, 609.25 μmol), BrettPhos Pd G3 ( 36.82 mg, 40.62 μmol), stir at 100°C for 2 hours. Water (10 mL) was added to the reaction solution and extracted with ethyl acetate (15 mL × 3). The organic phase was collected, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. It was then purified by column chromatography (dichloromethane: methanol = 10:0-10:1) to obtain compound 51c. MS ESI m/z:663.5[M+H] + .
步骤3 Step 3
在反应瓶中加入化合物51c(120.00mg,108.62μmol)，二氯甲烷(10mL)，三氟乙酸(2mL)，25℃搅拌1小时。氢氧化钠水溶液(1N)调节pH为8-9，用二氯甲烷萃取(10mL x 3)，无水硫酸钠干燥，过滤，减压浓缩得到化合物51d。Compound 51c (120.00 mg, 108.62 μmol), dichloromethane (10 mL), and trifluoroacetic acid (2 mL) were added to the reaction bottle, and stirred at 25°C for 1 hour. Adjust the pH to 8-9 with aqueous sodium hydroxide solution (1N), extract with dichloromethane (10mL x 3), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 51d.
步骤4Step 4
在反应瓶中加入化合物51d(100mg,177.71μmol)，甲醇(10mL)，甲醛(10.67mg,355.41μmol)，醋酸硼氢化钠(225.98mg,1.07mmol)，冰醋酸(32.01mg,533.12μmol)，25℃搅拌1小时。用氢氧化钠水溶液(1N)调节pH＝8-9，使用二氯甲烷萃取(20mL×2)，饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得到产物粗品。粗品经制备高效液相色谱分离(色谱柱:Phenomenex Gemini-NX 80×40mm×3μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:30-60％，8min)得到化合物51。MS ESI m/z:577.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.06(s,1H),8.69(s,1H),8.08(d,J＝4.00Hz,1H),7.41(d,J＝8.00Hz,1H),7.24-7.26(m,1H),6.46(d,J＝8.00Hz,1H),6.13(d,J＝4.00Hz,1H),4.73-4.86(m,3H),4.42(t,J＝8.00Hz,2H),4.11(t,J＝8.00Hz,2H),3.51-3.60(m,1H),3.45-3.50(m,2H),3.07-3.15(m,2H),2.74(d,J＝12.00Hz,2H),2.44(d,J＝8.00Hz,2H),2.21-2.30(m,5H),1.95-2.08(m,4H),1.72-1.83(m,2H),1.36(d,J＝8.00Hz,6H)。Add compound 51d (100 mg, 177.71 μmol), methanol (10 mL), formaldehyde (10.67 mg, 355.41 μmol), sodium acetate borohydride (225.98 mg, 1.07 mmol), glacial acetic acid (32.01 mg, 533.12 μmol) into the reaction bottle. Stir at 25°C for 1 hour. Adjust pH=8-9 with sodium hydroxide aqueous solution (1N), extract with dichloromethane (20 mL×2), wash with saturated brine (10 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX 80×40mm×3μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% ammonia bicarbonate); gradient: B%: 30-60 %, 8 min) to obtain compound 51. MS ESI m/z:577.4[M+H] + . 1H NMR (400MHz, CDCl3) δppm 9.06 (s, 1H), 8.69 (s, 1H), 8.08 (d, J = 4.00Hz, 1H), 7.41 (d, J = 8.00Hz, 1H), 7.24-7.26 ( m,1H),6.46(d,J＝8.00Hz,1H),6.13(d,J＝4.00Hz,1H),4.73-4.86(m,3H),4.42(t,J＝8.00Hz,2H), 4.11(t,J＝8.00Hz,2H),3.51-3.60(m,1H),3.45-3.50(m,2H),3.07-3.15(m,2H),2.74(d,J＝12.00Hz,2H) ,2.44(d,J＝8.00Hz,2H),2.21-2.30(m,5H),1.95-2.08(m,4H),1.72-1.83(m,2H),1.36(d,J＝8.00Hz,6H ).
实施例52
Example 52
步骤1step 1
将异丙醇(8mL)，4-氨基-2-氯嘧啶(0.3g,2.32mmol)，化合物52a(639.09mg,3.01mmol)加入长管中，开始搅拌；然后将三乙胺(1.17g,11.58mmol,1.61mL)加入其中，升温至100℃，反应20小时。向反应体系中加入20mL饱和氯化铵溶液后，加入乙酸乙酯萃取(15mL×2)萃取；得到的有机相用15mL饱和食盐水洗涤后，加入无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩，得到化合物52b。MS ESI m/z:306.3[M+H]+。1H NMR(400MHz,DMSO-d6)δppm 7.72(d,J＝6.0Hz,1H),6.41(s,2H),5.74(d,J＝5.6Hz,1H),4.27(d,J＝12.8Hz,2H),4.10-4.19(m,2H),2.84(d,J＝12.4Hz,2H),1.74-1.81(m,2H),1.51-1.59(m,2H),1.41(s,9H)。Add isopropanol (8mL), 4-amino-2-chloropyrimidine (0.3g, 2.32mmol), and compound 52a (639.09mg, 3.01mmol) into a long tube and start stirring; then triethylamine (1.17g, 11.58mmol, 1.61mL) was added, the temperature was raised to 100°C, and the reaction was carried out for 20 hours. After adding 20 mL saturated ammonium chloride solution to the reaction system, add ethyl acetate for extraction (15 mL Concentrate under reduced pressure to obtain compound 52b. MS ESI m/z:306.3[M+H] + . 1H NMR (400MHz, DMSO-d6) δppm 7.72 (d, J = 6.0Hz, 1H), 6.41 (s, 2H), 5.74 (d, J = 5.6Hz, 1H), 4.27 (d, J = 12.8Hz, 2H), 4.10-4.19 (m, 2H), 2.84 (d, J = 12.4Hz, 2H), 1.74-1.81 (m, 2H), 1.51-1.59 (m, 2H), 1.41 (s, 9H).
步骤2Step 2
在反应瓶中加入化合物41e(80mg,203.08μmol)，化合物52b(62.02mg,203.08μmol)，二氧六环(6mL)，氮气置换三次加入乙酸钾(59.79mg,609.25μmol)，BrettPhos Pd G3(36.82mg,40.62μmol)，100℃搅拌3小时。反应液中加入水(10mL)使用乙酸乙酯萃取(15mL×3)，收集有机相，有机相使用饱和食盐 水洗涤(20mL)，无水硫酸钠干燥，过滤，减压浓缩得到粗品，粗品通过柱层析纯化(二氯甲烷：甲醇＝10:0～10:1)纯化得到化合物52c。MS ESI m/z:663.2[M+H]+。Add compound 41e (80 mg, 203.08 μmol), compound 52b (62.02 mg, 203.08 μmol), dioxane (6 mL) to the reaction bottle, replace with nitrogen three times and add potassium acetate (59.79 mg, 609.25 μmol), BrettPhos Pd G3 ( 36.82 mg, 40.62 μmol), stir at 100°C for 3 hours. Add water (10 mL) to the reaction solution and extract with ethyl acetate (15 mL × 3). Collect the organic phase and use saturated salt for the organic phase. Wash with water (20 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product. The crude product is purified by column chromatography (dichloromethane:methanol=10:0~10:1) to obtain compound 52c. MS ESI m/z:663.2[M+H] + .
步骤3Step 3
在反应瓶中加入化合物52c(20mg,30.17μmol)，二氯甲烷(2mL)，三氟乙酸(0.4mL)25℃搅拌1小时。反应液使用1N氢氧化钠水溶液调节pH＝8-9，使用二氯甲烷萃取(10mL×3)，有机相使用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得到产物。无后续纯化，直接用于下一步反应。得到化合物52d。MS ESI m/z:563.4[M+H]+。Compound 52c (20 mg, 30.17 μmol), dichloromethane (2 mL), and trifluoroacetic acid (0.4 mL) were added to the reaction bottle and stirred at 25°C for 1 hour. The reaction solution was adjusted to pH=8-9 using 1N sodium hydroxide aqueous solution, extracted with dichloromethane (10 mL × 3), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the product . No subsequent purification was required and it was used directly in the next reaction. Compound 52d was obtained. MS ESI m/z:563.4[M+H] + .
步骤4Step 4
在反应瓶中加入化合物52d(20mg,35.54μmol)，甲醇(4mL)，甲醛(5.77mg,71.08μmol,5.29μL,37％纯度)，醋酸硼氢化钠(45.20mg,213.25μmol)，冰醋酸(6.40mg,106.62μmol,6.10μL)25℃搅拌1小时。反应液使用1N氢氧化钠水溶液调节pH＝8-9，使用二氯甲烷萃取(10mL×3)，收集有机相，有机相使用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得到产物。产物中加入甲醇(6mL)搅拌10min后过滤，滤饼减压浓缩得到产物。得到化合物52。MS ESI m/z:577.2[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.05(s,1H),8.68(s,1H),8.07(d,J＝8.00Hz,1H),7.41(d,J＝8.00Hz,1H),7.30(s,1H),6.46(d,J＝8.00Hz,1H),6.13(d,J＝4.00Hz,1H),4.71-4.80(m,1H),4.42(t,J＝8.00Hz,4H),4.05-4.13(m,2H),3.56(m,1H),3.43-3.50(m,2H),3.30(s,4H),3.05-3.16(m,2H),2.43(s,3H),2.20-2.30(m,2H),2.00-2.09(m,2H),1.69-1.80(m,4H),1.36(d,J＝8.00Hz,6H)。Add compound 52d (20 mg, 35.54 μmol), methanol (4 mL), formaldehyde (5.77 mg, 71.08 μmol, 5.29 μL, 37% purity), sodium acetate borohydride (45.20 mg, 213.25 μmol), glacial acetic acid ( 6.40 mg, 106.62 μmol, 6.10 μL) and stirred at 25°C for 1 hour. The reaction solution was adjusted to pH=8-9 using 1N sodium hydroxide aqueous solution, extracted with dichloromethane (10mL×3), and the organic phase was collected. The organic phase was washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and reduced to Concentrate under pressure to obtain the product. Methanol (6 mL) was added to the product and stirred for 10 min, then filtered, and the filter cake was concentrated under reduced pressure to obtain the product. Compound 52 was obtained. MS ESI m/z:577.2[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.05 (s, 1H), 8.68 (s, 1H), 8.07 (d, J = 8.00Hz, 1H), 7.41 (d, J = 8.00Hz, 1H), 7.30 ( s,1H),6.46(d,J＝8.00Hz,1H),6.13(d,J＝4.00Hz,1H),4.71-4.80(m,1H),4.42(t,J＝8.00Hz,4H), 4.05-4.13(m,2H),3.56(m,1H),3.43-3.50(m,2H),3.30(s,4H),3.05-3.16(m,2H),2.43(s,3H),2.20- 2.30(m,2H),2.00-2.09(m,2H),1.69-1.80(m,4H),1.36(d,J＝8.00Hz,6H).
实施例53
Example 53
步骤1step 1
将异丙醇(10mL)，4-氨基-2-氯嘧啶(0.5g,3.86mmol)，化合物53a(841.73mg,4.25mmol)依次加入反应瓶中，开始搅拌；然后再加入三乙胺(1.95g,19.30mmol,2.69mL)，升温至100℃，反应20小时。反应液在45℃下进行减压浓缩，加入乙酸乙酯(5mL)，搅拌20min，过滤，滤饼在40℃干燥浓缩得到化合物53b。MS ESI m/z:292.3[M+H]+。1H NMR(400MHz,CDCl3)δppm 7.70(d,J＝5.6Hz,1H),6.45(s,2H),5.73(d,J＝5.6Hz,1H),4.75-4.69(m,1H),4.39-4.23(m,2H),3.42-3.37(m,1H),3.33-3.28(m,1H),3.15-3.12(m,1H),1.84-1.81(m,2H),1.38(s,9H)。 Add isopropanol (10mL), 4-amino-2-chloropyrimidine (0.5g, 3.86mmol), and compound 53a (841.73mg, 4.25mmol) into the reaction bottle in sequence, and start stirring; then add triethylamine (1.95 g, 19.30mmol, 2.69mL), raise the temperature to 100°C, and react for 20 hours. The reaction solution was concentrated under reduced pressure at 45°C, ethyl acetate (5 mL) was added, stirred for 20 min, filtered, and the filter cake was dried and concentrated at 40°C to obtain compound 53b. MS ESI m/z:292.3[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 7.70 (d, J = 5.6 Hz, 1H), 6.45 (s, 2H), 5.73 (d, J = 5.6 Hz, 1H), 4.75-4.69 (m, 1H), 4.39-4.23(m,2H),3.42-3.37(m,1H),3.33-3.28(m,1H),3.15-3.12(m,1H),1.84-1.81(m,2H),1.38(s,9H ).
步骤2Step 2
将1,4-二氧六环(1.5mL)，化合物49c(10mg,25.45μmol)，化合物53b(7.41mg,25.45μmol)加入反应瓶中，搅拌；氮气置换，依次加入碳酸铯(24.88mg,76.35μmol)，BrettPhos Pd G3(4.61mg,5.09μmol)，升温至100℃，反应3小时。加入饱和氯化铵溶液(10mL)，用乙酸乙酯萃取(5mL×3)；合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液在45℃减压浓缩得粗品。通过硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～10:90)，得到化合物53c。MS ESI m/z:648.2[M+H]+。Add 1,4-dioxane (1.5 mL), compound 49c (10 mg, 25.45 μmol), and compound 53b (7.41 mg, 25.45 μmol) into the reaction flask, stir; replace with nitrogen, and add cesium carbonate (24.88 mg, 76.35 μmol), BrettPhos Pd G3 (4.61 mg, 5.09 μmol), raise the temperature to 100°C, and react for 3 hours. Add saturated ammonium chloride solution (10mL) and extract with ethyl acetate (5mL×3); combine the organic phases, wash with saturated brine (10mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 45°C to obtain Crude. Separation and purification by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0 to 10:90) gave compound 53c. MS ESI m/z:648.2[M+H] + .
步骤3Step 3
将二氯甲烷(1.5mL)，化合物53c(15mg,9.26μmol)加入反应瓶中，搅拌下，滴入三氟乙酸(4.04mmol,0.3mL)，室温20℃，反应1小时。反应液在40℃下减压浓缩，加入二氯甲烷(10mL)和饱和碳酸氢钠溶液(10mL)，分相；有机相依次用饱和碳酸氢钠溶液(10mL)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液在40℃下进行减压浓缩得到化合物53d。MS ESI m/z:548.2[M+H]+。Add dichloromethane (1.5 mL) and compound 53c (15 mg, 9.26 μmol) into the reaction flask, add trifluoroacetic acid (4.04 mmol, 0.3 mL) dropwise under stirring, and react at room temperature 20°C for 1 hour. The reaction solution was concentrated under reduced pressure at 40°C, dichloromethane (10 mL) and saturated sodium bicarbonate solution (10 mL) were added, and the phases were separated; the organic phase was washed with saturated sodium bicarbonate solution (10 mL) and saturated brine (10 mL). , dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40°C to obtain compound 53d. MS ESI m/z:548.2[M+H] + .
步骤4Step 4
将甲醇(1.5mL)，化合物53d(13.00mg,23.74μmol)，甲醛溶液(19.26mg,237.35μmol,37％纯度)加入反应瓶中，搅拌，室温20℃反应0.5小时；然后加入醋酸硼氢化钠(20.12mg,94.94μmol)，继续反应0.5小时。向反应体系中加入水(10mL)，饱和碳酸钠溶液(1.5mL)，用二氯甲烷萃取(10mL×2)；有机相依次采用饱和碳酸氢钠溶液(10mL)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液在40℃进行减压浓缩得粗品，经高效液相制备色谱分离纯化(色谱柱:Waters Xbridge BEH C18 100×30mm×5μm；流动相:A(乙腈)和B(水，含10mM碳酸氢铵)；梯度：B％:16％-45％，8min)，得到化合物53。MS ESI m/z:562.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.08(s,1H),8.62(s,1H),8.06(d,J＝5.6Hz,1H),7.41-7.37(m,2H),6.50(d,J＝8.0Hz,1H),6.22(d,J＝5.6Hz,1H),4.70(t,J＝7.6Hz,1H),4.24-4.21(m,1H),4.03-3.98(m,1H),3.89-3.79(m,2H),3.70-3.52(m,2H),3.33-3.26(m,2H),3.21-3.14(m,1H),3.07-2.99(m,1H),2.87-2.73(m,2H),2.62(s,3H),2.51-2.43(m,1H),2.32-2.24(m,2H),2.20-2.11(m,2H),1.94-1.89(m,2H),1.46(d,J＝6.0Hz,3H),1.37-1.35(m,6H)。Add methanol (1.5 mL), compound 53d (13.00 mg, 23.74 μmol), and formaldehyde solution (19.26 mg, 237.35 μmol, 37% purity) into the reaction bottle, stir, and react at room temperature 20°C for 0.5 hours; then add sodium acetate borohydride (20.12mg, 94.94μmol), continue the reaction for 0.5 hours. Add water (10 mL) and saturated sodium carbonate solution (1.5 mL) to the reaction system, and extract with dichloromethane (10 mL × 2); the organic phase is washed with saturated sodium bicarbonate solution (10 mL) and saturated brine (10 mL). , dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40°C to obtain a crude product, which was separated and purified by high-performance liquid phase preparative chromatography (chromatographic column: Waters Xbridge BEH C18 100 × 30 mm × 5 μm; mobile phase: A (acetonitrile) and B (water, containing 10mM ammonium bicarbonate); gradient: B%: 16%-45%, 8 min) to obtain compound 53. MS ESI m/z:562.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.08 (s, 1H), 8.62 (s, 1H), 8.06 (d, J = 5.6Hz, 1H), 7.41-7.37 (m, 2H), 6.50 (d, J ＝8.0Hz,1H),6.22(d,J＝5.6Hz,1H),4.70(t,J＝7.6Hz,1H),4.24-4.21(m,1H),4.03-3.98(m,1H),3.89 -3.79(m,2H),3.70-3.52(m,2H),3.33-3.26(m,2H),3.21-3.14(m,1H),3.07-2.99(m,1H),2.87-2.73(m, 2H),2.62(s,3H),2.51-2.43(m,1H),2.32-2.24(m,2H),2.20-2.11(m,2H),1.94-1.89(m,2H),1.46(d, J＝6.0Hz,3H),1.37-1.35(m,6H).
实施例54

Example 54

步骤1step 1
将1,4-二氧六环(3mL)，化合物49c(20.00mg,50.90μmol)，化合物52b(15.54mg,50.90μmol)加入反应瓶中，搅拌；氮气置换，依次将碳酸铯(49.75mg,152.69μmol)，BrettPhos Pd G3(6.92mg,7.63μmol)加入，升温至100℃，反应4小时。加入水(15mL)，用乙酸乙酯萃取(10mL×3)；合并有机相，饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，滤液于45℃减压浓缩。粗品经薄层色谱分离(展开剂为乙酸乙酯：甲醇＝20:1)，得到化合物54a。MS ESI m/z:662.5[M+H]+。Add 1,4-dioxane (3mL), compound 49c (20.00mg, 50.90μmol), and compound 52b (15.54mg, 50.90μmol) into the reaction flask, stir; replace with nitrogen, add cesium carbonate (49.75mg, 152.69 μmol), BrettPhos Pd G3 (6.92 mg, 7.63 μmol) was added, the temperature was raised to 100°C, and the reaction was carried out for 4 hours. Add water (15 mL) and extract with ethyl acetate (10 mL × 3); combine the organic phases, wash with saturated brine (20 mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 45°C. The crude product was separated by thin layer chromatography (developing solvent: ethyl acetate: methanol = 20:1) to obtain compound 54a. MS ESI m/z:662.5[M+H] + .
步骤2Step 2
将二氯甲烷(2mL)，化合物54a(10.00mg,15.11μmol)加入反应瓶中，搅拌；然后将三氟乙酸(614.00mg,5.39mmol)滴加入，室温20℃，反应1小时。加入水(10mL)和二氯甲烷(10mL)，用碳酸钠溶液(2M)将pH值调至8～9，分液；有机相依次用碳酸钠溶液(2M，10mL)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液在40℃减压浓缩得化合物54b。MS ESI m/z:562.4[M+H]+。Add dichloromethane (2 mL) and compound 54a (10.00 mg, 15.11 μmol) into the reaction bottle and stir; then add trifluoroacetic acid (614.00 mg, 5.39 mmol) dropwise, and react at room temperature 20°C for 1 hour. Add water (10 mL) and dichloromethane (10 mL), adjust the pH value to 8-9 with sodium carbonate solution (2M), and separate the liquids; use sodium carbonate solution (2M, 10 mL) and saturated saline solution (10 mL) for the organic phase in sequence. ), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40°C to obtain compound 54b. MS ESI m/z:562.4[M+H] + .
步骤3Step 3
将甲醇(2mL)，化合物54b(20.00mg,35.60μmol),甲醛溶液(28.89mg,356.04μmol,26.51μL,37％含量)加入反应瓶中，搅拌，室温20℃，反应0.5小时；加入三乙酰氧基硼氢化钠(30.18mg,142.41μmol)，继续反应0.5小时。依次加入水(10mL)，饱和碳酸钠溶液(1.5mL)，用二氯甲烷萃取(10mL×2)；合并的有机相依次用饱和碳酸氢钠溶液(10mL)，饱和食盐水(10mL)洗涤后，无水硫酸钠干燥，过滤，滤液于40℃下减压浓缩得粗品，经高效液相制备色谱分离(色谱柱:Waters Xbridge BEH C18 100×25mm×10μm；流动相:A(乙腈)和B(水，含10mM碳酸氢铵)；梯度：B％:15％-55％，8min)纯化得到化合物54。MS ESI m/z:576.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.08(s,1H),8.64(s,1H),8.07(d,J＝5.6Hz,1H),7.39(d,J＝8.0Hz,1H),7.31(s,1H),6.49(d,J＝8.0Hz,1H),6.15(d,J＝5.2Hz,1H),4.69(t,J＝7.6Hz,1H),4.44-4.41(m,2H),4.26-4.20(m,1H),3.80(t,J＝7.2Hz,1H),3.59-3.53(m,1H),3.41-3.25(m,5H),3.20-3.14(m,1H),3.07-2.99(m,1H),2.80-2.71(m,1H),2.50-2.46(m,4H),2.29-2.24(m,1H),2.18-2.05(m,3H),1.97-1.87(m,1H),1.78-1.76(m,2H),1.46(d,J＝6.0Hz,3H),1.36(d,J＝7.2Hz,6H)。Add methanol (2mL), compound 54b (20.00mg, 35.60μmol), formaldehyde solution (28.89mg, 356.04μmol, 26.51μL, 37% content) into the reaction bottle, stir, and react at room temperature 20°C for 0.5 hours; add triacetyl Sodium oxyborohydride (30.18 mg, 142.41 μmol), continue the reaction for 0.5 hours. Water (10 mL) and saturated sodium carbonate solution (1.5 mL) were added in sequence, and extracted with dichloromethane (10 mL × 2); the combined organic phase was washed with saturated sodium bicarbonate solution (10 mL) and saturated brine (10 mL). , dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40°C to obtain a crude product, which was separated by high-performance liquid phase preparative chromatography (chromatographic column: Waters Xbridge BEH C18 100 × 25 mm × 10 μm; mobile phase: A (acetonitrile) and B (Water, containing 10mM ammonium bicarbonate); gradient: B%: 15%-55%, 8min) was purified to obtain compound 54. MS ESI m/z:576.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.08 (s, 1H), 8.64 (s, 1H), 8.07 (d, J = 5.6Hz, 1H), 7.39 (d, J = 8.0Hz, 1H), 7.31 ( s,1H),6.49(d,J＝8.0Hz,1H),6.15(d,J＝5.2Hz,1H),4.69(t,J＝7.6Hz,1H),4.44-4.41(m,2H), 4.26-4.20(m,1H),3.80(t,J＝7.2Hz,1H),3.59-3.53(m,1H),3.41-3.25(m,5H),3.20-3.14(m,1H),3.07- 2.99(m,1H),2.80-2.71(m,1H),2.50-2.46(m,4H),2.29-2.24(m,1H),2.18-2.05(m,3H),1.97-1.87(m,1H ), 1.78-1.76 (m, 2H), 1.46 (d, J = 6.0Hz, 3H), 1.36 (d, J = 7.2Hz, 6H).
实施例55
Example 55
步骤1step 1
将1,4-二氧六环(3mL)，化合物49c(15mg,38.17μmol)，化合物51b(11.66mg,38.17μmol)加入反应瓶中，搅拌；氮气置换，依次加入碳酸铯(37.31mg,114.52μmol)，BrettPhos Pd G3(5.19mg,5.73μmol)，升温至100℃，反应4小时。降至室温，加水(15mL)，用乙酸乙酯萃取(10mL×3)；合并有机相，饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，滤液于45℃减压浓缩得粗品，经薄层色谱法(TLC)分离(展开剂：乙酸乙酯：甲醇＝20:1)，得到化合物55a。MS ESI m/z:662.3[M+H]+。Add 1,4-dioxane (3mL), compound 49c (15mg, 38.17μmol), and compound 51b (11.66mg, 38.17μmol) into the reaction flask, stir; replace with nitrogen, add cesium carbonate (37.31mg, 114.52 μmol), BrettPhos Pd G3 (5.19 mg, 5.73 μmol), raise the temperature to 100°C, and react for 4 hours. Cool to room temperature, add water (15 mL), and extract with ethyl acetate (10 mL × 3); combine the organic phases, wash with saturated brine (20 mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 45°C to obtain a crude product. After separation by thin layer chromatography (TLC) (developing solvent: ethyl acetate: methanol = 20:1), compound 55a was obtained. MS ESI m/z:662.3[M+H] + .
步骤2Step 2
将二氯甲烷(2mL)，化合物55a(15mg,22.66μmol)加入反应瓶中，搅拌；滴入三氟乙酸(614.00mg,5.39mmol,0.4mL)，于18℃反应1小时。加入水(10mL)和二氯甲烷(10mL)，用碳酸钠溶液(2M)将pH值调至8～9，分液；有机相依次用碳酸钠溶液(2M)溶液(10mL)，饱和食盐水(10mL)洗涤，有机相用无水硫酸钠干燥，过滤，滤液于40℃减压浓缩得到化合物55b。MS ESI m/z:562.3[M+H]+。Add dichloromethane (2 mL) and compound 55a (15 mg, 22.66 μmol) into the reaction flask, stir; add trifluoroacetic acid (614.00 mg, 5.39 mmol, 0.4 mL) dropwise, and react at 18°C for 1 hour. Add water (10 mL) and dichloromethane (10 mL), adjust the pH value to 8~9 with sodium carbonate solution (2M), and separate the liquids; use sodium carbonate solution (2M) solution (10 mL) and saturated saline solution for the organic phase in turn. (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40°C to obtain compound 55b. MS ESI m/z:562.3[M+H] + .
步骤3Step 3
将甲醇(2mL)，化合物55b(15mg,22.66μmol)加入反应瓶中，搅拌；滴加甲醛溶液(18.39mg,226.64μmol,37％含量)，于18℃反应0.5小时；加入三乙酰氧基硼氢化钠(19.21mg,90.65μmol)，反应0.5小时。加入水(10mL)，饱和碳酸钠溶液(1.5mL)，用二氯甲烷萃取(10mL×2)；合并有机相，依次用饱和碳酸氢钠溶液(10mL)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液于40℃减压浓缩得到粗品。加入甲醇(0.5mL)溶解，滴加水(0.5mL)，搅拌5分钟，过滤得到化合物55。MS ESI m/z:576.5[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.08(s,1H),8.67(s,1H),8.07(d,J＝5.6Hz,1H),7.39(d,J＝8.0Hz,1H),7.32(s,1H),6.49(d,J＝7.6Hz,1H),6.16-6.12(m,1H),4.90-4.82(m,2H),4.70(t,J＝7.6Hz,1H),4.26-4.20(m,1H),3.82-3.79(m,1H),3.58-3.51(m,1H),3.33-3.26(m,1H),3.20-3.14(m,1H),3.07-2.99(m,1H),2.80-2.73(m,3H),2.50-2.43(m,3H),2.30-2.27(m,4H),2.20-2.10(m,1H),2.08-1.97(m,4H),1.95-1.87(m,1H),1.46(d,J＝6.0Hz,3H),1.36(t,J＝6.8Hz,6H)。 Add methanol (2 mL) and compound 55b (15 mg, 22.66 μmol) into the reaction bottle, stir; add formaldehyde solution (18.39 mg, 226.64 μmol, 37% content) dropwise, and react at 18°C for 0.5 hours; add triacetoxyboron Sodium hydride (19.21 mg, 90.65 μmol), react for 0.5 hours. Add water (10mL), saturated sodium carbonate solution (1.5mL), and extract with dichloromethane (10mL×2); combine the organic phases, wash with saturated sodium bicarbonate solution (10mL), saturated brine (10mL), and wash without Dry over sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 40°C to obtain crude product. Methanol (0.5 mL) was added to dissolve, water (0.5 mL) was added dropwise, stirred for 5 minutes, and filtered to obtain compound 55. MS ESI m/z:576.5[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.08 (s, 1H), 8.67 (s, 1H), 8.07 (d, J = 5.6Hz, 1H), 7.39 (d, J = 8.0Hz, 1H), 7.32 ( s,1H),6.49(d,J＝7.6Hz,1H),6.16-6.12(m,1H),4.90-4.82(m,2H),4.70(t,J＝7.6Hz,1H),4.26-4.20 (m,1H),3.82-3.79(m,1H),3.58-3.51(m,1H),3.33-3.26(m,1H),3.20-3.14(m,1H),3.07-2.99(m,1H) ,2.80-2.73(m,3H),2.50-2.43(m,3H),2.30-2.27(m,4H),2.20-2.10(m,1H),2.08-1.97(m,4H),1.95-1.87( m, 1H), 1.46 (d, J = 6.0Hz, 3H), 1.36 (t, J = 6.8Hz, 6H).
实施例56
Example 56
步骤1step 1
将二氧六环(3mL)，化合物49c(20.00mg,50.90μmol)，化合物48b(14.83mg,50.90μmol)加入反应瓶中，搅拌；氮气置换，依次加入碳酸铯(49.75mg,152.69μmol)，BrettPhos Pd G3(9.23mg,10.18μmol)，升温至100℃，反应3小时。加水(15mL)，用乙酸乙酯萃取(10mL×3)；合并有机相，饱和食盐水洗涤(20mL)，无水硫酸钠干燥，过滤，滤液于45℃减压浓缩得粗品，粗品经薄层色谱法(TLC)分离得到化合物56a。MS ESI m/z:648.5[M+H]+。Add dioxane (3mL), compound 49c (20.00mg, 50.90μmol), and compound 48b (14.83mg, 50.90μmol) into the reaction flask, stir; replace with nitrogen, add cesium carbonate (49.75mg, 152.69μmol) in sequence, BrettPhos Pd G3 (9.23 mg, 10.18 μmol), heated to 100°C, and reacted for 3 hours. Add water (15 mL) and extract with ethyl acetate (10 mL Compound 56a was isolated by chromatography (TLC). MS ESI m/z:648.5[M+H] + .
步骤2Step 2
将化合物56a(10mg,15.44μmol),二氯甲烷(2mL)，三氟乙酸(0.4mL)加入反应瓶中，于25℃搅拌1小时。将反应液滴入到氢氧化钠水溶液(1N)中，调节pH＝8-9，用二氯甲烷萃取(10mL×3)，合并有机相。用饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，减压浓缩得化合物56b。MS ESI m/z:548.4[M+H]+。Compound 56a (10 mg, 15.44 μmol), dichloromethane (2 mL), and trifluoroacetic acid (0.4 mL) were added to the reaction flask, and stirred at 25°C for 1 hour. The reaction solution was dropped into sodium hydroxide aqueous solution (1N), adjusted to pH = 8-9, extracted with dichloromethane (10 mL × 3), and the organic phases were combined. Wash with saturated brine (10 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 56b. MS ESI m/z:548.4[M+H] + .
步骤3Step 3
将化合物56b(10mg,18.26μmol),甲醇(4mL)，甲醛(1.10mg,36.52μmol)，醋酸硼氢化钠(23.22mg,109.55μmol)，冰醋酸(3.29mg,54.77μmol)依次加入反应瓶中，25℃搅拌1小时。加入氢氧化钠水溶液(1N)，调节pH＝7～8，用二氯甲烷萃取(10mL×3)，合并有机相，饱和食盐水洗涤(10mL)，无水硫酸钠干燥，过滤，滤液经Pre-HPLC分离(色谱柱:Waters Xbridge Prep OBD C18 150×40mm×10μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢铵)；梯度：B％:30-60％，8min)得到化合物56。MS ESI m/z:548.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.09(s,1H),8.82(s,1H),8.13(s,1H),7.34-7.48(m,2H),6.50(d,J＝8.00Hz,1H),6.20(s,1H),5.31(s,1H),4.71(m,1H),4.23(s,1H),4.10(m,1H),3.92(m,2H),3.82(m,3H),3.55(m,1H),3.29(d,J＝8.00Hz,1H),3.16(m,1H),3.00-3.10(m,1H),2.73-2.88(m,2H),2.47(m,1H),2.28-2.38(m,4H),2.13-2.20 (m,1H),1.90-1.95(m,1H),1.71(d,J＝8.00Hz,1H),1.46(d,J＝4.00Hz,3H),1.34(b,6H)。Add compound 56b (10 mg, 18.26 μmol), methanol (4 mL), formaldehyde (1.10 mg, 36.52 μmol), sodium acetate borohydride (23.22 mg, 109.55 μmol), and glacial acetic acid (3.29 mg, 54.77 μmol) into the reaction bottle in sequence. , stir at 25°C for 1 hour. Add sodium hydroxide aqueous solution (1N), adjust the pH to 7~8, extract with dichloromethane (10mL -HPLC separation (column: Waters Xbridge Prep OBD C18 150×40mm×10μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% ammonium bicarbonate); gradient: B%: 30-60%, 8min) Compound 56 was obtained. MS ESI m/z:548.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.09 (s, 1H), 8.82 (s, 1H), 8.13 (s, 1H), 7.34-7.48 (m, 2H), 6.50 (d, J = 8.00Hz, 1H ),6.20(s,1H),5.31(s,1H),4.71(m,1H),4.23(s,1H),4.10(m,1H),3.92(m,2H),3.82(m,3H) ,3.55(m,1H),3.29(d,J＝8.00Hz,1H),3.16(m,1H),3.00-3.10(m,1H),2.73-2.88(m,2H),2.47(m,1H ),2.28-2.38(m,4H),2.13-2.20 (m,1H),1.90-1.95(m,1H),1.71(d,J＝8.00Hz,1H),1.46(d,J＝4.00Hz,3H),1.34(b,6H).
实施例57
Example 57
步骤1step 1
将异丙醇(6mL)，化合物19g(0.6g,1.81mmol)加入反应瓶中，搅拌；加入氨水(2.4mL,28％纯度)，升温至70℃，反应3小时。加入20mL水，用乙酸乙酯(15mL×2)萃取；合并有机相，饱和食盐水(15mL)洗涤，无水硫酸钠干燥，过滤，滤液于45℃减压浓缩得到化合物57a。MS ESI m/z:253.4[M+H]+。Add isopropyl alcohol (6 mL) and compound 19g (0.6 g, 1.81 mmol) into the reaction flask and stir; add ammonia water (2.4 mL, 28% purity), raise the temperature to 70°C, and react for 3 hours. Add 20 mL of water and extract with ethyl acetate (15 mL × 2); combine the organic phases, wash with saturated brine (15 mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 45°C to obtain compound 57a. MS ESI m/z:253.4[M+H] + .
步骤2Step 2
将二氯甲烷(15mL)，化合物57a(450mg,1.78mmol)加入反应瓶中，搅拌；加入三乙胺(451.11mg,4.46mmol)，温度降至0～10℃，滴加3-氯丙磺酰氯(410.43mg,2.32mmol)，升至20℃，反应0.5小时。加入饱和氯化铵溶液(20mL)，分液，有机相用无水硫酸钠干燥后，过滤，滤液于40℃减压浓缩得粗品，粗品经快速硅胶柱层析纯化(石油醚:乙酸乙酯＝100:0～60:40)得到化合物57b。MS ESI m/z:393.4[M+H]+。Add dichloromethane (15 mL) and compound 57a (450 mg, 1.78 mmol) into the reaction flask and stir; add triethylamine (451.11 mg, 4.46 mmol), lower the temperature to 0~10°C, and add 3-chloropropanesulfonate dropwise. Acid chloride (410.43 mg, 2.32 mmol), raised to 20°C, reacted for 0.5 hours. Add saturated ammonium chloride solution (20 mL), separate the layers, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 40°C to obtain a crude product, which is purified by flash silica gel column chromatography (petroleum ether: ethyl acetate). =100:0～60:40) to obtain compound 57b. MS ESI m/z:393.4[M+H] + .
步骤3Step 3
将N,N-二甲基甲酰胺(9mL)，化合物57b(450mg,1.15mmol)加入反应瓶中，搅拌下加入1.8-二氮杂二环[5.4.0]十一烷-7-烯(523.04mg,3.44mmol)，于20℃下反应5小时。加入饱和氯化铵溶液(25mL)，乙酸乙酯萃取(15mL×2)，合并有机相，饱和食盐(20mL)水洗涤，无水硫酸钠干燥，过滤，滤液于45℃下减压浓缩得粗品，经快速硅胶柱层析纯化(石油醚:乙酸乙酯＝100：0～50：50)得到物化合物57c。1H NMR(400MHz,DMSO-d6)δppm 7.47-7.35(m,4H),7.31-7.28(m,2H),7.25-7.18(m,4H),4.38(s,1H),3.60-3.52(m,2H),3.43-3.36(m,1H),3.24-3.13(m,4H),2.89-2.86(m,1H),2.37-2.30(m,2H),0.82-0.81(m,3H)。Add N,N-dimethylformamide (9mL) and compound 57b (450mg, 1.15mmol) into the reaction flask, and add 1.8-diazabicyclo[5.4.0]undecane-7-ene ( 523.04 mg, 3.44 mmol), reacted at 20°C for 5 hours. Add saturated ammonium chloride solution (25 mL), extract with ethyl acetate (15 mL , purified by flash silica gel column chromatography (petroleum ether:ethyl acetate=100:0~50:50) to obtain compound 57c. 1 H NMR (400MHz, DMSO-d 6 ) δppm 7.47-7.35(m,4H),7.31-7.28(m,2H),7.25-7.18(m,4H),4.38(s,1H),3.60-3.52( m,2H),3.43-3.36(m,1H),3.24-3.13(m,4H),2.89-2.86(m,1H),2.37-2.30(m,2H),0.82-0.81(m,3H).
步骤4Step 4
将甲醇(10mL)，氢氧化钯/碳(196.98mg,280.52μmol,20％纯度),化合物57c(0.5g,1.40mmol)依次加入反应瓶中，搅拌下，加入盐酸(12M,233.77μL)，氢气(15Psi)下于20℃，反应20小时。过滤，滤液在45℃下进行减压浓缩得到化合物57d的盐酸盐。MS ESI m/z:190.9[M+H]+。Methanol (10 mL), palladium hydroxide on carbon (196.98 mg, 280.52 μmol, 20% purity), and compound 57c (0.5 g, 1.40 mmol) were added to the reaction flask in sequence. Under stirring, hydrochloric acid (12 M, 233.77 μL) was added. React for 20 hours under hydrogen gas (15Psi) at 20°C. Filter, and the filtrate is concentrated under reduced pressure at 45°C to obtain the hydrochloride of compound 57d. MS ESI m/z:190.9[M+H] + .
步骤5Step 5
将1,4二氧六环(5mL)，化合物15g(70mg,245.98μmol)，化合物57d的盐酸盐(60.84mg,319.77μmol)加入反应瓶中，开始搅拌；氮气置换，依次加入碳酸铯(320.58mg,983.91μmol)，Xantphos Pd G4(35.51mg,36.90μmol)，温度升至100℃后，反应2小时。加入饱和氯化铵溶液(15mL)，乙酸乙酯萃取(10mL×2)，合并有机相，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液在45℃下减压浓缩 得粗品，粗品经快速硅胶柱纯化(石油醚:乙酸乙酯＝100:0～30:70)化合物57e。MS ESI m/z:394.3[M+H]+。Add 1,4 dioxane (5mL), compound 15g (70mg, 245.98μmol), and the hydrochloride of compound 57d (60.84mg, 319.77μmol) into the reaction flask, start stirring; replace with nitrogen, and add cesium carbonate ( 320.58 mg, 983.91 μmol), Xantphos Pd G4 (35.51 mg, 36.90 μmol). After the temperature was raised to 100°C, the reaction was carried out for 2 hours. Add saturated ammonium chloride solution (15mL), extract with ethyl acetate (10mL×2), combine the organic phases, wash with saturated brine (10mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 45°C. The crude product was obtained, and the crude product was purified by fast silica gel column (petroleum ether:ethyl acetate=100:0~30:70) compound 57e. MS ESI m/z:394.3[M+H] + .
步骤6Step 6
将1,4二氧六环(5mL)，化合物57e(60mg,152.31μmol)，化合物23f(31.01mg,137.08μmol)加入反应瓶中，搅拌；置换氮气，依次加入碳酸铯(124.07mg,380.78μmol)，BrettPhos Pd G3(20.71mg,22.85μmol)，升温至100℃，反应2小时。加入饱和氯化铵溶液(15mL)，乙酸乙酯萃取(10mL×2)，合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液在45℃下进行减压浓缩得粗品，经制备薄层色谱法分离(展开剂为乙酸乙酯：甲醇＝40:1，)，得到化合物57。MS ESI m/z:584.5[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.09(s,1H),8.58(s,1H),8.08(d,J＝6.0Hz,1H),7.53-7.42(m,2H),6.59(d,J＝7.6Hz,1H),6.26-6.17(m,1H),4.92-4.77(m,1H),4.64-4.57(m,2H),4.50-4.29(m,2H),4.07-4.02(m,1H),3.90-3.80(m,2H),3.67-3.57(m,3H),3.52(s,3H),3.35-3.32(m,2H),3.23-3.19(m,2H),2.43-2.36(m,2H),2.10-2.02(m,1H),1.93-1.81(m,1H),1.53(d,J＝6.4Hz,3H),1.39-1.36(m,6H)。Add 1,4 dioxane (5mL), compound 57e (60mg, 152.31μmol), and compound 23f (31.01mg, 137.08μmol) into the reaction flask, stir; replace nitrogen, and add cesium carbonate (124.07mg, 380.78μmol) in sequence ), BrettPhos Pd G3 (20.71 mg, 22.85 μmol), heated to 100°C, and reacted for 2 hours. Add saturated ammonium chloride solution (15mL), extract with ethyl acetate (10mL×2), combine the organic phases, wash with saturated brine (10mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure at 45°C. The crude product was obtained and separated by preparative thin layer chromatography (the developing solvent was ethyl acetate: methanol = 40:1,) to obtain compound 57. MS ESI m/z:584.5[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.09 (s, 1H), 8.58 (s, 1H), 8.08 (d, J = 6.0Hz, 1H), 7.53-7.42 (m, 2H), 6.59 (d, J ＝7.6Hz,1H),6.26-6.17(m,1H),4.92-4.77(m,1H),4.64-4.57(m,2H),4.50-4.29(m,2H),4.07-4.02(m,1H ),3.90-3.80(m,2H),3.67-3.57(m,3H),3.52(s,3H),3.35-3.32(m,2H),3.23-3.19(m,2H),2.43-2.36(m ,2H),2.10-2.02(m,1H),1.93-1.81(m,1H),1.53(d,J=6.4Hz,3H),1.39-1.36(m,6H).
实施例58
Example 58
步骤1step 1
将化合物49c(50.00mg,127.25μmol)，化合物45a(28.16mg,127.25μmol)，二氧六环(4mL)依次加入反应瓶中，氮气置换三次，加入乙酸钾(37.46mg,381.74μmol)，BrettPhos Pd G3(23.07mg,25.45μmol)，100℃搅拌4小时。反应液过滤，收集滤液，减压浓缩得粗品经高效液相色谱分离(色谱柱:Phenomenex Luna C18 75×30mm×3μm；流动相:A(乙腈)和B(水，含0.04％盐酸)；梯度：B％:15-50％)得到化合物58。MS ESI m/z:578.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.09(s,1H),8.44(s,1H),8.09(d,J＝4.00Hz,1H),7.40(d,J＝8.00Hz,1H),7.36(s,1H),6.50(d,J＝8.00Hz,1H),6.29(d,J＝4.00Hz,1H),5.09(d,J＝12.00Hz,2H),4.71(t,J＝8.00Hz,1H),4.23(t,J＝4.00Hz,1H),3.81(t,J＝8.00Hz,1H),3.53(m,1H),3.25-3.34(m,1H),3.18(m,2H),2.91-3.08(m,3H),2.77(m,1H),2.66(s,6H),2.47(m,1H),2.28(m,1H),2.21(m,3H),1.90-1.97(m,1H),1.70(m,2H),1.47(d,J＝4.00Hz,3H),1.38(t,J＝8.00Hz,6H)。Add compound 49c (50.00 mg, 127.25 μmol), compound 45a (28.16 mg, 127.25 μmol), and dioxane (4 mL) into the reaction bottle in sequence, replace with nitrogen three times, add potassium acetate (37.46 mg, 381.74 μmol), and BrettPhos Pd G3 (23.07 mg, 25.45 μmol), stirred at 100°C for 4 hours. The reaction solution was filtered, the filtrate was collected, and the crude product was concentrated under reduced pressure and separated by high performance liquid chromatography (chromatographic column: Phenomenex Luna C18 75 × 30 mm × 3 μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% hydrochloric acid); gradient :B%:15-50%) to obtain compound 58. MS ESI m/z:578.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.09 (s, 1H), 8.44 (s, 1H), 8.09 (d, J = 4.00Hz, 1H), 7.40 (d, J = 8.00Hz, 1H), 7.36 ( s,1H),6.50(d,J＝8.00Hz,1H),6.29(d,J＝4.00Hz,1H),5.09(d,J＝12.00Hz,2H),4.71(t,J＝8.00Hz, 1H),4.23(t,J＝4.00Hz,1H),3.81(t,J＝8.00Hz,1H),3.53(m,1H),3.25-3.34(m,1H),3.18(m,2H), 2.91-3.08(m,3H),2.77(m,1H),2.66(s,6H),2.47(m,1H),2.28(m,1H),2.21(m,3H),1.90-1.97(m, 1H), 1.70 (m, 2H), 1.47 (d, J = 4.00Hz, 3H), 1.38 (t, J = 8.00Hz, 6H).
实施例59

Example 59

步骤1step 1
将乙醇(40mL)，化合物59a(2.0g,8.15mmol)加入反应瓶中，搅拌；依次加入乙酸钠(2.01g,24.46mmol)，羟胺盐酸盐(1.70g,24.46mmol)，于20℃反应2小时。加入饱和碳酸氢钠溶液(80mL)，用乙酸乙酯萃取(30mL×2)；合并有机相，用30mL饱和食盐水洗涤，无水硫酸钠干燥，过滤，45℃减压浓缩得到化合物59b。1H NMR(400MHz,CDCl3)δppm 7.47-7.46(m,1H),7.38-7.32(m,4H),5.19-5.08(m,2H),4.55(d,J＝33.2Hz,1H),3.52-3.47(m,1H),3.39-3.33(m,1H),3.20(s,1H),2.66-2.52(m,1H),2.45-2.38(m,1H),1.93-1.87(m,1H),1.75-1.72(m,1H)。Add ethanol (40mL) and compound 59a (2.0g, 8.15mmol) into the reaction flask and stir; add sodium acetate (2.01g, 24.46mmol) and hydroxylamine hydrochloride (1.70g, 24.46mmol) in sequence, and react at 20°C 2 hours. Add saturated sodium bicarbonate solution (80 mL) and extract with ethyl acetate (30 mL × 2); combine the organic phases, wash with 30 mL saturated brine, dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure at 45°C to obtain compound 59b. 1 H NMR (400MHz, CDCl 3 ) δppm 7.47-7.46 (m, 1H), 7.38-7.32 (m, 4H), 5.19-5.08 (m, 2H), 4.55 (d, J = 33.2Hz, 1H), 3.52 -3.47(m,1H),3.39-3.33(m,1H),3.20(s,1H),2.66-2.52(m,1H),2.45-2.38(m,1H),1.93-1.87(m,1H) ,1.75-1.72(m,1H).
步骤2Step 2
将丙酮(30mL)，化合物59b(1.6g,6.15mmol)加反应瓶中，搅拌；然后加入对甲苯磺酸酰氯(1.76g,9.22mmol)，20℃搅拌10分钟；然后将碳酸钠(1.95g,18.44mmol)溶于水(30mL)，滴加到反应液中，继续反应22小时。加入水(50mL)，用乙酸乙酯萃取(30mL×2)；合并有机相，饱和碳酸氢钠溶液(30mL)洗涤，无水硫酸钠干燥，过滤，滤液于45℃进行减压浓缩得化合物59c。Add acetone (30mL) and compound 59b (1.6g, 6.15mmol) into the reaction flask and stir; then add p-toluenesulfonic acid chloride (1.76g, 9.22mmol) and stir at 20°C for 10 minutes; then add sodium carbonate (1.95g ,18.44mmol) was dissolved in water (30mL), added dropwise to the reaction solution, and the reaction was continued for 22 hours. Add water (50 mL) and extract with ethyl acetate (30 mL .
步骤3Step 3
将丙酮(20mL)，化合物59c(1.5g,3.62mmol)加入反应瓶中，搅拌；加入浓盐酸(12M,753.97μL)于20℃反应22小时。加入水(35mL)，用饱和碳酸钠溶液将pH值调至8～9，用乙酸乙酯萃取(20mL×4)；合并有机相，饱和食盐水(40mL)洗涤，无水硫酸钠干燥，过滤，滤液在45℃减压浓缩得粗品，粗品经过硅胶柱层析分离纯化(梯度洗脱：石油醚:乙酸乙酯＝100:0～0:1)得到化合物59d。MS ESI m/z:261.2[M+H]+；1H NMR(400MHz,CDCl3)δppm 7.39-7.33(m,5H),6.73-6.66(m,1H),5.31-5.08(m,2H),4.45-4.36(m,1H),3.97-3.90(m,1H),3.70-3.63(m,1H),3.50-3.43(m,1H),2.87-2.69(m,1H),2.52-2.44(m,1H),2.11-2.04(m,1H),2.00-1.97(m,1H)。Add acetone (20 mL) and compound 59c (1.5 g, 3.62 mmol) into the reaction bottle and stir; add concentrated hydrochloric acid (12 M, 753.97 μL) and react at 20°C for 22 hours. Add water (35 mL), adjust the pH to 8-9 with saturated sodium carbonate solution, and extract with ethyl acetate (20 mL × 4); combine the organic phases, wash with saturated brine (40 mL), dry over anhydrous sodium sulfate, and filter. , the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product, which was separated and purified by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 100:0~0:1) to obtain compound 59d. MS ESI m/z:261.2[M+H] + ; 1 H NMR (400MHz, CDCl 3 )δppm 7.39-7.33(m,5H),6.73-6.66(m,1H),5.31-5.08(m,2H) ,4.45-4.36(m,1H),3.97-3.90(m,1H),3.70-3.63(m,1H),3.50-3.43(m,1H),2.87-2.69(m,1H),2.52-2.44( m,1H),2.11-2.04(m,1H),2.00-1.97(m,1H).
步骤4Step 4
将四氢呋喃(10mL)，化合物59d(0.2g,768.38μmol)加入反应瓶中，搅拌；氮气置换，将温度降至0～5℃，滴加硼烷二甲基硫醚络合物(10M,768.38μL)，升温至20℃，反应15小时，温度降至0～5℃，将甲醇(5mL)缓慢滴加到其中，搅拌15分钟后，反应液在45℃下进行减压浓缩得粗品，加入甲苯(10mL)溶解后，再加入丙烷(1.26mL)，升温至60℃，反应3小时。缓慢加入甲醇(10mL)，搅拌15min，然后进行减压浓缩，加入饱和碳酸氢钠溶液(20mL)，用二氯甲烷萃取(15mL×2)，合并有机相，无水硫酸钠干燥，过滤，滤液在40℃下减压浓缩得粗品，粗品通过硅胶柱层析分离(梯度洗脱：石油醚:乙酸乙酯＝1:1～0:1，二氯甲烷：甲醇＝80:1～15:1(含1％氨水，体积比))得到化合物59e。MS ESI m/z:247.2[M+H]+。1H NMR(400MHz,CDCl3)δ＝7.40-7.31(m,5H),5.15-5.17(m,2H),4.37-4.27(m,1H),3.66-3.64(m,1H),3.58-3.52(m,2H),3.07-2.92(m,2H),1.93-1.82(m,4H),1.68-1.60(m,1H)。 Add tetrahydrofuran (10 mL) and compound 59d (0.2 g, 768.38 μmol) into the reaction flask and stir; replace with nitrogen, lower the temperature to 0~5°C, and add borane dimethyl sulfide complex (10 M, 768.38 μmol) dropwise. μL), raise the temperature to 20°C, and react for 15 hours. The temperature drops to 0~5°C. Methanol (5mL) is slowly added dropwise to it. After stirring for 15 minutes, the reaction solution is concentrated under reduced pressure at 45°C to obtain a crude product. Add After toluene (10 mL) was dissolved, propane (1.26 mL) was added, the temperature was raised to 60°C, and the reaction was carried out for 3 hours. Methanol (10 mL) was slowly added, stirred for 15 min, then concentrated under reduced pressure, added saturated sodium bicarbonate solution (20 mL), extracted with dichloromethane (15 mL × 2), combined the organic phases, dried over anhydrous sodium sulfate, filtered, and the filtrate Concentrate under reduced pressure at 40°C to obtain a crude product, which is separated by silica gel column chromatography (gradient elution: petroleum ether: ethyl acetate = 1:1 ~ 0:1, dichloromethane: methanol = 80:1 ~ 15:1 (containing 1% ammonia water, volume ratio)) to obtain compound 59e. MS ESI m/z:247.2[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δ = 7.40-7.31 (m, 5H), 5.15-5.17 (m, 2H), 4.37-4.27 (m, 1H), 3.66-3.64 (m, 1H), 3.58-3.52 (m,2H),3.07-2.92(m,2H),1.93-1.82(m,4H),1.68-1.60(m,1H).
步骤5Step 5
将化合物59e(200mg,812.00μmol)，4-氨基-2-氯嘧啶(105.19mg,812.00μmol)，异丙醇(8mL)，三乙胺(493.00mg,4.87mmol,678.13μL)，依次加入反应瓶中，于100℃搅拌12小时。降至25℃，减压浓缩得到粗品，粗品经快速柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)得到化合物59f。MS ESI m/z:340.3[M+H]+。Compound 59e (200 mg, 812.00 μmol), 4-amino-2-chloropyrimidine (105.19 mg, 812.00 μmol), isopropyl alcohol (8 mL), and triethylamine (493.00 mg, 4.87 mmol, 678.13 μL) were added to the reaction in sequence. bottle and stirred at 100°C for 12 hours. The mixture was lowered to 25°C and concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (dichloromethane:methanol=10:0-10:1) to obtain compound 59f. MS ESI m/z:340.3[M+H] + .
步骤6Step 6
将化合物41e(100mg,253.85μmol)，化合物59f(86.16mg,253.85μmol)，乙酸钾(74.74mg,761.56μmol)，二氧六环(6mL)依次加入反应瓶中，氮气置换三次，加入BrettPhos Pd G3(46.02mg,50.77μmol)，100℃搅拌2小时。降温至25℃，加入水(10mL)，乙酸乙酯萃取(10mL×3)，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩得到粗品，粗品经快速硅胶柱层析纯化(二氯甲烷：甲醇＝10:0-10:1)得到化合物59g。MS ESI m/z:697.4[M+H]+。Add compound 41e (100 mg, 253.85 μmol), compound 59f (86.16 mg, 253.85 μmol), potassium acetate (74.74 mg, 761.56 μmol), and dioxane (6 mL) into the reaction flask in sequence, replace with nitrogen three times, and add BrettPhos Pd G3 (46.02 mg, 50.77 μmol), stirred at 100°C for 2 hours. Cool to 25°C, add water (10 mL), extract with ethyl acetate (10 mL Methyl chloride: methanol = 10:0-10:1) to obtain 59 g of compound. MS ESI m/z:697.4[M+H] + .
步骤7Step 7
将化合物59g(50mg,71.75μmol)，三乙胺(7.26mg,71.75μmol,9.99μL)，三乙基硅烷(16.69mg,143.50μmol,22.92μL)，二氯甲烷(6mL)依次加入反应瓶中，氮气置换三次，加入氯化钯(12.72mg,71.75μmol)，Add compound 59g (50mg, 71.75μmol), triethylamine (7.26mg, 71.75μmol, 9.99μL), triethylsilane (16.69mg, 143.50μmol, 22.92μL) and dichloromethane (6mL) into the reaction bottle in sequence , nitrogen was replaced three times, and palladium chloride (12.72 mg, 71.75 μmol) was added.
25℃搅拌2小时。减压浓缩得到化合物59h。MS ESI m/z:563.4[M+H]+。Stir at 25°C for 2 hours. Concentrate under reduced pressure to obtain compound 59h. MS ESI m/z:563.4[M+H] + .
步骤9Step 9
将化合物59h(40mg,71.08μmol)，冰醋酸(12.81mg,213.25μmol,12.21μL)，醋酸硼氢化钠(90.39mg,426.49μmol)，甲醇(5mL)，甲醛(4.27mg,142.16μmol,3.92μL)依次加入反应瓶中，氮气置换三次，25℃搅拌1小时。过滤，收集滤液，减压浓缩得粗品，粗品经高效液相色谱分离(色谱柱:Waters Xbridge Prep OBD C18 150×40mm×10μm；流动相:A(乙腈)和B(水，含0.04％碳酸氢氨)；梯度：B％:30-60％)化合物59。MS ESI m/z:577.4[M+H]+。1H NMR(400MHz,CDCl3)δppm 9.05(s,1H),8.66(s,1H),8.07(d,J＝4.00Hz,1H),7.41(d,J＝8.00Hz,1H),6.46(d,J＝8.00Hz,1H),6.11(d,J＝8.00Hz,1H),5.60(s,1H),4.75(m,1H),4.56-4.66(m,1H),4.42(t,J＝8.00Hz,2H),4.10(t,J＝8.00Hz,2H),3.52-3.64(m,1H),3.43-3.50(m,2H),3.23-3.40(m,2H),3.08-3.14(m,2H),2.97-3.07(m,2H),2.61(s,3H),2.21-2.30(m,2H),1.88-2.04(m,2H),1.74-1.77(m,2H),1.56-1.65(m,3H),1.36(dd,J＝4.00,4.00Hz,6H)。Compound 59h (40 mg, 71.08 μmol), glacial acetic acid (12.81 mg, 213.25 μmol, 12.21 μL), sodium borohydride acetate (90.39 mg, 426.49 μmol), methanol (5 mL), formaldehyde (4.27 mg, 142.16 μmol, 3.92 μL) ) were added into the reaction bottle in turn, replaced with nitrogen three times, and stirred at 25°C for 1 hour. Filter, collect the filtrate, and concentrate under reduced pressure to obtain a crude product, which is separated by high performance liquid chromatography (chromatographic column: Waters Xbridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: A (acetonitrile) and B (water, containing 0.04% hydrogen carbonate) Ammonia); Gradient: B%: 30-60%) Compound 59. MS ESI m/z:577.4[M+H] + . 1 H NMR (400MHz, CDCl 3 ) δppm 9.05 (s, 1H), 8.66 (s, 1H), 8.07 (d, J = 4.00Hz, 1H), 7.41 (d, J = 8.00Hz, 1H), 6.46 ( d,J＝8.00Hz,1H),6.11(d,J＝8.00Hz,1H),5.60(s,1H),4.75(m,1H),4.56-4.66(m,1H),4.42(t,J ＝8.00Hz,2H),4.10(t,J＝8.00Hz,2H),3.52-3.64(m,1H),3.43-3.50(m,2H),3.23-3.40(m,2H),3.08-3.14( m,2H),2.97-3.07(m,2H),2.61(s,3H),2.21-2.30(m,2H),1.88-2.04(m,2H),1.74-1.77(m,2H),1.56- 1.65(m,3H),1.36(dd,J＝4.00,4.00Hz,6H).
生物测试数据：Biological test data:
测试例1：化合物对EGFR(T790M/C797S/L858R)、EGFR(Del19/C797S/L858R)、EGFR野生型的激酶抑制活性Test Example 1: Kinase inhibitory activity of compounds against EGFR (T790M/C797S/L858R), EGFR (Del19/C797S/L858R), and EGFR wild type
在反应缓冲液(20mM羟乙基哌嗪乙硫磺酸(Hepes)(pH 7.5),10mM氯化镁(MgCl2),1mM乙二醇双氨乙基醚四乙酸(EGTA),0.02％聚氧乙烯十二烷醚(Brij35),0.02mg/mL BSA,0.1mM钒酸钠(Na3VO4),2mM二硫苏糖醇(DTT),1％DMSO)中依次加入一定浓度的底物、辅酶因子、激酶和测试化合物(10个剂量，3倍连续稀释液，2％DMSO最终浓度)并混匀，将混合物在室温下孵育20分钟，向反应混合液中加入一定浓度的33P-ATP开始反应，随后室温孵育120分钟。最后通过过滤-结合的方法来检测反应物的放射性。最终的激酶活性以测试样品中剩余的激酶活性占DMSO对照组的激酶活性的比例来表示。通过GraphPad软件拟合量效关系曲线并计算IC50。In reaction buffer (20mM hydroxyethyl piperazine ethyl sulfonic acid (Hepes) (pH 7.5), 10mM magnesium chloride (MgCl 2 ), 1mM ethylene glycol bisaminoethyl ether tetraacetic acid (EGTA), 0.02% polyoxyethylene ten Dialkyl ether (Brij35), 0.02mg/mL BSA, 0.1mM sodium vanadate (Na 3 VO 4 ), 2mM dithiothreitol (DTT), 1% DMSO) and then add a certain concentration of substrate and coenzyme factor , kinase and test compound (10 doses, 3-fold serial dilution, 2% DMSO final concentration) and mix well, incubate the mixture at room temperature for 20 minutes, add a certain concentration of 33 P-ATP to the reaction mixture to start the reaction , followed by incubation at room temperature for 120 minutes. Finally, the radioactivity of the reactants is detected by filtration-binding method. The final kinase activity is expressed as the ratio of the remaining kinase activity in the test sample to the kinase activity in the DMSO control group. The dose-response relationship curve was fitted and IC 50 was calculated using GraphPad software.
实验结果如表1所示。The experimental results are shown in Table 1.
表1本发明化合物EGFR激酶的抑制活性IC50测试结果

Table 1 IC 50 test results of the inhibitory activity of EGFR kinase of the compounds of the present invention

注：“/”代表未测试Note: "/" means not tested
实验结论：本发明的化合物对EGFR(T790M/C797S/L858R)、EGFR(Del19/C797S/L858R)激酶有很强的抑制活性，对EGFR野生型激酶抑制活性弱。Experimental conclusion: The compound of the present invention has strong inhibitory activity against EGFR (T790M/C797S/L858R) and EGFR (Del19/C797S/L858R) kinases, but has weak inhibitory activity against EGFR wild-type kinase.
测试例2：化合物对Ba/F3-FL-EGFR(T790M/C797S/L858R)、EGFR(DeL19/C797S/L858R)、Ba/F3-FL-EGFR细胞增殖的抑制活性Test Example 2: Inhibitory activity of compounds on Ba/F3-FL-EGFR (T790M/C797S/L858R), EGFR (DeL19/C797S/L858R), and Ba/F3-FL-EGFR cell proliferation
三磷酸腺苷(Adenosine Tri-Phosphate,ATP)是自然界中各种生命活动中共用的能量载体，是能量储存和转移的最小单位。CellTiter-GloTM活细胞检测试剂盒采用萤光素酶作检测物，发光过程中萤光素酶需要ATP的参与。向细胞培养基中加入CellTiter-GloTM试剂，测量发光值，光信号和体系中ATP量成正比，而ATP又和活细胞数正相关。因此通过使用CellTiter-Glo试剂盒检测ATP含量，可以检测出细胞的增殖情况。本测试中，细胞系为Ba/F3-FL-EGFR(T790M/C797S/L858R)、Ba/F3-FL-EGFR(DeL19-T790M-C797S)、Ba/F3-FL-EGFR稳转细胞株。Adenosine Tri-Phosphate (ATP) is a common energy carrier in various life activities in nature and is the smallest unit of energy storage and transfer. CellTiter-Glo TM live cell detection kit uses luciferase as the detection substance, and luciferase requires the participation of ATP during the luminescence process. Add CellTiter-Glo TM reagent to the cell culture medium and measure the luminescence value. The light signal is directly proportional to the amount of ATP in the system, and ATP is positively correlated with the number of viable cells. Therefore, by using the CellTiter-Glo kit to detect ATP content, cell proliferation can be detected. In this test, the cell lines were Ba/F3-FL-EGFR (T790M/C797S/L858R), Ba/F3-FL-EGFR (DeL19-T790M-C797S), and Ba/F3-FL-EGFR stably transfected cell lines.
IC50测定过程：IC 50 determination process:
1)细胞培养1) Cell culture
将细胞系在培养条件37℃，5％CO2的培养箱中进行培养。定期传代，取处于对数生长期的细胞用于铺板。The cell lines were cultured in an incubator with culture conditions of 37°C and 5% CO2 . Passage regularly and use cells in the logarithmic growth phase for plating.
2)化合物存储板制备2) Compound storage plate preparation
a)用DMSO将待测化合物配置成10mM溶液，再用DMSO将待测化合物稀释至0.3或1mM。a) Use DMSO to prepare the compound to be tested into a 10mM solution, and then use DMSO to dilute the compound to be tested to 0.3 or 1mM.
b)制备1000×化合物存储板(管)：用DMSO从最高浓度3倍梯度稀释至最低浓度，9个浓度。b) Prepare 1000× compound storage plate (tube): use DMSO to dilute 3-fold gradient from the highest concentration to the lowest concentration, 9 concentrations.
c)20×化合物工作液的配制:在平底的96孔透明药板中加入49μL细胞培养液，从1000×化合物存储板中吸取1μL化合物加入96孔透明药板的细胞培养液中。在溶媒对照中加入1μL DMSO。加入化合物或DMSO后用排枪吹打混匀。c) Preparation of 20× compound working solution: Add 49 μL of cell culture medium to a flat-bottomed 96-well transparent medicine plate, pipet 1 μL of compound from the 1000× compound storage plate and add it to the cell culture medium of the 96-well transparent medicine plate. Add 1 μL DMSO to the vehicle control. Add the compound or DMSO and mix with pipette.
3)细胞铺板与给药3) Cell plating and drug administration
a)用台盼兰进行细胞染色并计数活细胞，要求细胞活率90％以上。a) Use trypan blue to stain cells and count viable cells. The cell viability rate is required to be above 90%.
b)在化合物检测细胞板中每孔加入95μL细胞悬液(2000细胞数/孔)，在Min对照孔中加入不含细胞(含0.1％DMSO)的培养液。b) Add 95 μL of cell suspension (2000 cells/well) to each well of the compound detection cell plate, and add culture medium without cells (containing 0.1% DMSO) to the Min control well.
c)化合物检测细胞板加药：取5μL的20×化合物工作液按表1所示加入到细胞培养板中。在Max对照中加入5μL DMSO-细胞培养液混合液。DMSO终浓度为0.1％。c) Compound detection cell plate addition: Take 5 μL of 20× compound working solution and add it to the cell culture plate as shown in Table 1. Add 5 μL of DMSO-cell culture medium mixture to the Max control. The final DMSO concentration is 0.1%.
d)将培养板在37℃,5％CO 2培养箱中培养72小时。d) Incubate the culture plate in a 37°C, 5% CO 2 incubator for 72 hours.
4)CellTiter-Glo发光法细胞活性检测4)CellTiter-Glo luminescence method cell viability detection
以下步骤按照Promega CellTiter-Glo发光法细胞活性检测试剂盒(Promega-G7573)的说明书来进行。The following steps are carried out according to the instructions of Promega CellTiter-Glo Luminescence Cell Viability Detection Kit (Promega-G7573).
a)将CellTiter-Glo缓冲液融化并放置至室温。 a) Melt CellTiter-Glo buffer and bring to room temperature.
b)将CellTiter-Glo底物放置至室温。b) Let CellTiter-Glo substrate come to room temperature.
c)在一瓶CellTiter-Glo底物中加入CellTiter-Glo缓冲液以溶解底物，从而配制CellTiter-Glo工作液。c) Add CellTiter-Glo buffer to a bottle of CellTiter-Glo substrate to dissolve the substrate to prepare CellTiter-Glo working solution.
d)缓慢涡旋震荡使充分溶解。d) Vortex slowly to fully dissolve.
e)取出细胞培养板放置10分钟使其平衡至室温。e) Take out the cell culture plate and let it equilibrate to room temperature for 10 minutes.
f)在每孔中加入50μL(等于每孔中细胞培养液一半体积)的CellTiter-Glo工作液。f) Add 50 μL (equal to half the volume of cell culture medium in each well) of CellTiter-Glo working solution into each well.
g)将培养板在轨道摇床上振摇2分钟以诱导细胞裂解。g) Shake the plate on an orbital shaker for 2 minutes to induce cell lysis.
h)培养板在室温放置10分钟以稳定发光信号。h) Place the culture plate at room temperature for 10 minutes to stabilize the luminescence signal.
i)在PerkinElmer2105读板器上检测发光信号。i) At PerkinElmer The luminescence signal is detected on the 2105 plate reader.
5)数据处理5)Data processing
PerkinElmer2105读数得出对应的每孔荧光值RLU。PerkinElmer 2105 readings yield the corresponding fluorescence value RLU for each well.
细胞增殖抑制率(Inhibition Rate)数据采用下列公式来处理：Cell proliferation inhibition rate (Inhibition Rate) data is processed using the following formula:
Inhibition Rate(Inh％)＝100-(RLUDrug-RLUMin)/(RLUMax-RLUMin)*100％。Inhibition Rate(Inh%)=100-(RLUDrug-RLUMin)/(RLUMax-RLUMin)*100%.
在EXCEL中计算不同浓度化合物对应的抑制率，然后用GraphPad Prism软件作抑制率曲线图并计算IC50值。Calculate the inhibition rates corresponding to different concentrations of compounds in EXCEL, and then use GraphPad Prism software to draw the inhibition rate curve and calculate the IC 50 value.
实验结果如表2所示。The experimental results are shown in Table 2.
表2本发明化合物EGFR突变型及野生型细胞增殖的抑制活性IC50测试结果
Table 2 IC 50 test results of the inhibitory activity of EGFR mutant and wild-type cell proliferation of the compounds of the present invention
注：“/”代表未测试。Note: "/" means not tested.
实验结论：本发明的化合物对EGFR三突变细胞具有较好增殖抑制活性，对EGFR野生型增殖抑制活性显著弱于突变细胞。Experimental conclusion: The compound of the present invention has good proliferation inhibitory activity on EGFR triple mutant cells, and its proliferation inhibitory activity on EGFR wild-type cells is significantly weaker than that of mutant cells.
测试例3：小鼠体内药代动力学评价Test Example 3: Pharmacokinetic evaluation in mice
实验过程：将2％DMSO+10％PEG400+88％水溶液经尾静脉注射到雌性Balb/c小鼠体内(过夜禁食，7-9周龄)。将2％DMSO+10％PEG400+88％水溶液灌胃给予雌性Balb/c小鼠(过夜禁食，7-9周龄)，给药剂量见表3。两组动物分别于给药后0.0833、0.25、0.5、1.0、2.0、4.0、8.0、24h从颈静脉和0.25、0.5、 1.0、2.0、4.0、6.0、8.0、24h从尾静脉采血约30μL置于添加了EDTA-K2的抗凝管中，离心分离血浆。采用LC-MS/MS法测定血药浓度，使用WinNonlinTM Version 6.3(Pharsight,Mountain View,CA)药动学软件，以非房室模型线性对数梯形法计算相关药代动力学参数。Experimental process: 2% DMSO + 10% PEG400 + 88% aqueous solution was injected into female Balb/c mice through the tail vein (overnight fasting, 7-9 weeks old). The 2% DMSO + 10% PEG400 + 88% aqueous solution was administered intragastrically to female Balb/c mice (overnight fasting, 7-9 weeks old). The dosage is shown in Table 3. The animals in the two groups were taken from the jugular vein and 0.25, 0.5, 24h after administration respectively. At 1.0, 2.0, 4.0, 6.0, 8.0, and 24 hours, approximately 30 μL of blood was collected from the tail vein and placed in anticoagulant tubes added with EDTA-K2, and the plasma was separated by centrifugation. The blood drug concentration was measured using the LC-MS/MS method, and WinNonlin TM Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software was used to calculate relevant pharmacokinetic parameters using the noncompartmental model linear logarithmic trapezoidal method.
实验结果如表3所示：The experimental results are shown in Table 3:
表3.药代动力学数据
Table 3. Pharmacokinetic data
注：“-”表示该参数无法通过计算得到；C0代表初始浓度；Cmax代表达峰浓度；T1/2代表消除半衰期；Vdss代表稳态表观分布容积；Cl代表总清除率；AUC0-last代表从0时间到最后一个可定量时间点的血浆浓度-时间曲线下面积；F(％)代表生物利用度，采用AUC0-last计算。Note: “-” means that the parameter cannot be calculated; C 0 represents the initial concentration; C max represents the peak concentration; T 1/2 represents the elimination half-life; V dss represents the steady-state apparent distribution volume; Cl represents the total clearance rate; AUC 0-last represents the area under the plasma concentration-time curve from time 0 to the last quantifiable time point; F (%) represents bioavailability, calculated using AUC 0-last .
本发明化合物16，27和32均展示良好的体内代谢稳定性，静脉给药清除率为16，58和40mL/min/kg，口服吸收生物利用度分别为63％，36％，69％。Compounds 16, 27 and 32 of the present invention all exhibit good in vivo metabolic stability, with intravenous clearance rates of 16, 58 and 40 mL/min/kg, and oral absorption bioavailability of 63%, 36% and 69% respectively.
结论：本发明化合物具有良好的体内代谢稳定性，优异的口服吸收药物暴露量和良好的口服吸收生物利用度。Conclusion: The compound of the present invention has good in vivo metabolic stability, excellent oral absorption drug exposure and good oral absorption bioavailability.
测试例4：本发明化合物的体外肝微粒体稳定性Test Example 4: In vitro liver microsome stability of the compounds of the present invention
通过人和小鼠肝微粒体代谢稳定性考察发明化合物在人上体外的代谢稳定性。试验条件：在37℃条件下，1μM的化合物分别与人肝微粒体辅以NADPH再生体系孵育一定的时间最长达60分钟后，采用LC-MS/MS方法检测所产生的样品中化合物的浓度。The metabolic stability of the inventive compound in vitro in humans was investigated through the metabolic stability of human and mouse liver microsomes. Test conditions: At 37°C, 1 μM of the compound was incubated with human liver microsomes supplemented with NADPH regeneration system for a certain period of time up to 60 minutes, and then the LC-MS/MS method was used to detect the concentration of the compound in the resulting sample. .
试验过程：96孔孵育板，分别命名为T0、T5、T15、T30、T45、T60、Blank60和NCF60。孵育板对应的反应时间点分别为0、5、15、30、45和60分钟。Blank60板中不加入供试品或对照化合物并在孵育60分钟后取样。NCF60板中用磷酸钾盐缓冲液代替NADPH再生体系溶液进行孵育60分钟。所有时间点样品均为单孔。Test process: 96-well incubation plates, named T0, T5, T15, T30, T45, T60, Blank60 and NCF60 respectively. The reaction time points corresponding to the incubation plates were 0, 5, 15, 30, 45 and 60 minutes respectively. Blank60 plates are filled with no test or control compounds and samples are taken after 60 minutes of incubation. The NCF60 plate was incubated with potassium phosphate buffer instead of NADPH regeneration system solution for 60 minutes. All time point samples are single wells.
分别在T0、T5、T15、T30、T45、T60和NCF60板上添加5μL供试品或对照品工作液和100μL微粒体工作液(肝微粒体蛋白浓度为0.5mg/mL)，在Blank60板中只添加微粒体工作液，然后将除T0和NCF60外的孵育板Blank60、T5、T15、T30、T45和T60放置于37℃水浴锅中预孵育大约10分钟。Add 5 μL of test or reference working solution and 100 μL of microsomal working solution (liver microsomal protein concentration is 0.5 mg/mL) to T0, T5, T15, T30, T45, T60 and NCF60 plates respectively. In the Blank60 plate Only add the microsomal working solution, and then place the incubation plates Blank60, T5, T15, T30, T45 and T60 except T0 and NCF60 in a 37°C water bath for pre-incubation for about 10 minutes.
T0板样品先加入180μL的终止液(终止液是含100ng/mL甲苯磺丁脲的乙腈：甲醇(95:5,V/V)溶液)后再添加NADPH再生体系工作液。To the T0 plate sample, first add 180 μL of stop solution (the stop solution is an acetonitrile:methanol (95:5, V/V) solution containing 100 ng/mL tolbutamide), and then add the NADPH regeneration system working solution.
在NCF60板上每孔添加50μL磷酸钾盐缓冲液，孵育60分钟。Add 50 μL potassium phosphate buffer to each well of the NCF60 plate and incubate for 60 minutes.
孵育板Blank60、T5、T15、T30、T45和T60预孵育结束后，每个样品孔内添加44μL NADPH再生体系 工作液以启动反应。因此，对于含供试品或对照品工作液的样品孔，反应终浓度为1μM，肝微粒体的浓度为0.5mg/mL，DMSO和乙腈在反应体系中的终浓度分别为0.01％(v/v)和0.99％(v/v)。After the pre-incubation of incubation plates Blank60, T5, T15, T30, T45 and T60, add 44μL NADPH regeneration system to each sample well. working fluid to initiate the reaction. Therefore, for the sample wells containing the test or reference working solution, the final concentration of the reaction is 1 μM, the concentration of liver microsomes is 0.5 mg/mL, and the final concentrations of DMSO and acetonitrile in the reaction system are 0.01% (v/ v) and 0.99% (v/v).
孵育适当时间(如5、15、30、45和60分钟)后，分别在Blank60、T5、T15、T30、T45、T60和NCF60板的每个供试品样品孔和对照品样品孔中加入180μL的终止液以终止反应。After incubating for an appropriate time (such as 5, 15, 30, 45 and 60 minutes), add 180 μL to each test sample well and control sample well of Blank60, T5, T15, T30, T45, T60 and NCF60 plates respectively. stop solution to stop the reaction.
所有样品板摇匀并离心10分钟，分别取80μL供试品上清液稀释到240μL乙腈：水(1:9,V/V)含0.1％甲酸的溶液中用于LC-MS/MS分析。All sample plates were shaken and centrifuged for 10 minutes. 80 μL of the test sample supernatant was diluted into 240 μL of acetonitrile:water (1:9, V/V) solution containing 0.1% formic acid for LC-MS/MS analysis.
通过对相应的各时间点与化合物的剩余百分比进行计算，得到化合物在人肝微粒体代谢的半衰期T1/2和清除率CLint(liver)。实验结果如表4所示。




By calculating the remaining percentage of the compound at each corresponding time point, the half-life T 1/2 and clearance rate CL int(liver) of the compound in human liver microsomal metabolism were obtained. The experimental results are shown in Table 4.




表4本发明化合物体外人肝微粒体代谢稳定性研究结果
Table 4 Research results on the in vitro human liver microsome metabolic stability of the compounds of the present invention
结论：本发明化合物在人和小鼠肝微粒体上具有良好的稳定性。 Conclusion: The compounds of the present invention have good stability on human and mouse liver microsomes.

Claims (21)

  1. 式(XI)所示化合物、其立体异构体或其药学上可接受的盐，
    The compound represented by formula (XI), its stereoisomer or its pharmaceutically acceptable salt,
    其中，in,
    T1和T2分别独立地选自CH和N；T 1 and T 2 are independently selected from CH and N;
    T4选自C和N；T 4 is selected from C and N;
    T5选自CH和N；T 5 is selected from CH and N;
    环A选自吡咯基、吡唑基、苯基、吡啶基、嘧啶基、吡啶酮基、嘧啶酮基、四氢吡啶基、四氢吡嗪基、二氢吡嗪酮基、2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；Ring A is selected from the group consisting of pyrrolyl, pyrazolyl, phenyl, pyridyl, pyrimidinyl, pyridonyl, pyrimidinonyl, tetrahydropyridinyl, tetrahydropyrazinyl, dihydropyrazinonyl, 2,3- dihydrobenzofuryl, phthalyl, 2,3-dihydropyridofuryl and isoindolinyl;
    环B选自吡唑基、哌啶基、四氢吡啶基、二氢吡喃基、环己基、环己烯基和7-10元杂环烷基；Ring B is selected from pyrazolyl, piperidinyl, tetrahydropyridinyl, dihydropyranyl, cyclohexyl, cyclohexenyl and 7-10 membered heterocycloalkyl;
    R1选自-C(Ra)2S(＝O)2C(Rb)3、 R 1 is selected from -C(R a ) 2 S(=O) 2 C(R b ) 3 ,
    R2选自H、卤素和OH，或者不存在；R 2 is selected from H, halogen and OH, or is absent;
    或者，T4选自C，R1和R2形成双键，使结构单元形成 Alternatively, T 4 is selected from C, R 1 and R 2 form a double bond such that the structural unit form
    或者，R1和R2与相连的原子成环形成四氢吡喃基；Alternatively, R 1 and R 2 form a ring with the connected atom to form a tetrahydropyranyl group;
    R3选自H、卤素、C1-3烷基、C1-3烷氧基和环丙基，所述C1-3烷基、C1-3烷氧基和环丙基分别独立地任选被1、2或3个卤素取代；R 3 is selected from H, halogen, C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl, the C 1-3 alkyl, C 1-3 alkoxy and cyclopropyl are independently optionally substituted by 1, 2 or 3 halogens;
    R4选自F、OH、C1-3烷基、C1-3烷氧基、和C1-3烷氨基，所述C1-3烷基、C1-3烷氧基和C1-3烷氨基分别独立地任选被1、2或3个卤素取代；R 4 is selected from F, OH, C 1-3 alkyl, C 1-3 alkoxy, and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino groups are each independently optionally substituted by 1, 2 or 3 halogens;
    R5选自＝CRaS(＝O)2CH3、 R 5 is selected from =CR a S(=O) 2 CH 3 ,
    R6和R7分别独立地选自H、卤素和C(Ra)3；R 6 and R 7 are each independently selected from H, halogen and C(R a ) 3 ;
    或者，R6和R7形成双键，使结构单元形成 Alternatively, R 6 and R 7 form a double bond, making the structural unit form
    或者，R6和R7与相连的原子成环形成四氢吡喃基；Alternatively, R 6 and R 7 form a ring with the connected atom to form a tetrahydropyranyl group;
    R8选自H和卤素； R 8 is selected from H and halogen;
    T3选自C(Rc)2、NRc和O；T 3 is selected from C(R c ) 2 , NR c and O;
    m选自0、1、2和3；m is selected from 0, 1, 2 and 3;
    n选自1、2和3；n is selected from 1, 2 and 3;
    r选自1、2和3；r is selected from 1, 2 and 3;
    s选自1和2；s is selected from 1 and 2;
    t选自1和2；t is selected from 1 and 2;
    各Ra分别独立地选自H和卤素；Each R a is independently selected from H and halogen;
    各Rb分别独立地选自H和卤素；Each R b is independently selected from H and halogen;
    Rc选自H和C1-3烷基；R c is selected from H and C 1-3 alkyl;
    条件是，requirement is,
    1)当R1选自-C(Ra)2S(＝O)2C(Rb)3时，环A选自2,3-二氢苯并呋喃基、苯酞基、2,3-二氢吡啶并呋喃基和异吲哚啉基；1) When R 1 is selected from -C(R a ) 2 S(=O) 2 C(R b ) 3 , ring A is selected from 2,3-dihydrobenzofuryl, phthalyl, 2,3 -Dihydropyridofuranyl and isoindolinyl;
    2)当R1选自时，环B不选自吡唑基。2) When R 1 is selected from When , Ring B is not selected from pyrazolyl.
  2. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，T3选自CH2、NH、NCH3、NCH2CH3、NCH(CH3)2和O。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein T 3 is selected from CH 2 , NH, NCH 3 , NCH 2 CH 3 , NCH(CH 3 ) 2 and O.
  3. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R1选自-CH2S(＝O)2CH3、-CHFS(＝O)2CH3、-CF2S(＝O)2CH3、 The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from -CH 2 S(=O) 2 CH 3 , -CHFS(=O) 2 CH 3 , -CF 2 S(=O) 2 CH 3 ,
  4. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R2选自H、F和OH，或者不存在。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R2 is selected from H, F and OH, or is absent.
  5. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R5选自＝CHS(＝O)2CH3、 The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 5 is selected from =CHS(=O) 2 CH 3 ,
  6. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R6和R7分别独立地选自H和CH3。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 6 and R 7 are independently selected from H and CH 3 .
  7. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，结构单元选自 The compound according to claim 1, its stereoisomer or its pharmaceutically acceptable salt, wherein the structural unit Selected from
  8. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，结构单元选自 The compound according to claim 1, its stereoisomer or its pharmaceutically acceptable salt, wherein the structural unit Selected from
  9. 根据权利要求1或8所述化合物、其立体异构体或其药学上可接受的盐，其中，结构单元 选自 The compound according to claim 1 or 8, its stereoisomer or its pharmaceutically acceptable salt, wherein the structural unit Selected from
  10. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R3选自F、CH3、CH2CH3、CH(CH3)2、OCH3和环丙基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from F, CH 3 , CH 2 CH 3 , CH(CH 3 ) 2 , OCH 3 and cyclopropane base.
  11. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，结构单元选自 The compound according to claim 1, its stereoisomer or its pharmaceutically acceptable salt, wherein the structural unit Selected from
  12. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R8选自H和F。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 8 is selected from H and F.
  13. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，环B选自 The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein ring B is selected from
  14. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R4选自F、OH、CH3、OCH3、CH2CH3、N(CH3)2和 The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 4 is selected from F, OH, CH 3 , OCH 3 , CH 2 CH 3 , N(CH 3 ) 2 and
  15. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，结构单元选自 The compound according to claim 1, its stereoisomer or its pharmaceutically acceptable salt, wherein the structural unit Selected from
  16. 根据权利要求15所述化合物、其立体异构体或其药学上可接受的盐，其中，结构单元选自 The compound according to claim 15, its stereoisomer or its pharmaceutically acceptable salt, wherein the structural unit Selected from
  17. 下列化合物、其立体异构体或其药学上可接受的盐，










    The following compounds, their stereoisomers or their pharmaceutically acceptable salts,










  18. 根据权利要求17所述化合物、其立体异构体或其药学上可接受的盐，其化合物选自，




































    The compound according to claim 17, its stereoisomer or its pharmaceutically acceptable salt, the compound is selected from,




































  19. 一种药物组合物，包括治疗有效量的根据权利要求1-17任意一项所述的化合物、其立体异构体或其药学上可接受的盐作为活性成分以及药学上可接受的载体、稀释剂或赋形剂。A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1-17, its stereoisomer or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier, diluent agents or excipients.
  20. 根据权利要求1-17任意一项所述化合物、其立体异构体或其药学上可接受的盐或者权利要求18所述组合物在制备治疗实体瘤药物中的应用。The use of the compound according to any one of claims 1 to 17, its stereoisomer or a pharmaceutically acceptable salt thereof or the composition according to claim 18 in the preparation of a drug for treating solid tumors.
  21. 根据权利要求19所述应用，其中实体瘤指非小细胞肺癌等实体瘤。 The application according to claim 19, wherein the solid tumor refers to solid tumors such as non-small cell lung cancer.
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WO2024152986A1 (en) * 2023-01-18 2024-07-25 北京鞍石生物科技有限责任公司 Heteroaryl amino compound, preparation method therefor, and use thereof

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